Reduction of Sterols and Other Compounds from Oils

ABSTRACT

A process for the removal of sterols, specifically cholesterol, from a triglyceride oil, preferably a marine triglyceride oil, said process comprising contacting an oil with an absorbent, specifically TRIS YL™, clay or a mixture thereof, heating the mixture to 100° C. to 210° C., preferably 150° C. to 170° C., preferably for a time period of greater than one minute and optionally at a pressure less than 1.33 Pa, preferably less than 1.33 Pa.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of priority to U.S. ProvisionalApplication No. 60/712,029, filed on Aug. 26, 2005, which isincorporated by reference herein in its entirety.

FIELD

The disclosed matter relates to methods of removing and/or reducingcompounds from oils (e.g., marine oils) and to oils prepared by thedisclosed methods. Methods of bleaching oils are also disclosed.

BACKGROUND

Polyunsaturated fatty acids (PUPA) such as the omega-3 fatty acids arevital to everyday life and function. For example, the beneficial effectsof omega-3 fatty acids like cis-5,8,11,14,17-eicosapentacnoic acid (EPA)and cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) on lowering serumtriglycerides are now well established. These compounds are also knownfor other cardioprotective benefits. See e.g., Dyrberg, et al, In: ω-3Fatty Acids: Prevention and Treatment of Vascular Disease. Kristensen,et al., eds., Bi & Gi Publ., Verona-Springer-Verlag, London, pp. 217-26,1995; O'Keefe and Harris, Am. J. Cardiology 2000, 85:1239-41; Radack etal., “The effects of low doses of omega-3 fatty acid supplementation onblood pressure in hypertensive subjects: a randomized controlled trial.”Arch. Intern. Med. 1991, 151:1173-1180. Indeed, the American HeartAssociation has also reported that omega-3 fatty acids can reducecardiovascular and heart disease risk. Other benefits of PUFAs are thoserelated to the prevention and/or treatment of inflammation,neurogenerative diseases, and cognitive development. See e.g., Sugano,Michihiro, “Balanced intake of polyunsaturated fatty acids for healthbenefits.” J. Oleo Sci. 2001, 50(5):305-311.

A primary source of many polyunsaturated fatty acids, including omega-3fatty acids, is fish oil. Diets rich in fish oils are known to have manybeneficial effects for heart disease, cancer, arthritis, allergies, andother chronic diseases. (See e.g., The American Heart Association,Scientific Statement, “Fish Consumption, Fish Oil, Omega-3 Fatty Acidsand Cardiovascular Disease,” November 2002; Appel et al., “Doessupplementation of diet with ‘fish oil’ reduce blood pressure? Ameta-analysis of controlled clinical trials.” Arch. Intern. Med. 1993,153(12):1429-1438; GISSI-Prevenzione Investigators. “Dietarysupplementation with omega-3 polyunsaturated fatty acids and vitamin Bafter myocardial infarction: results of the GASSI-Prevenzione trial.”Lancet 1999, 354:447-455.)

Some fish oils can contain compounds that may be undesirable for aparticular purpose. As such, various methods have been tried in order toremove and/or reduce undesirable compounds from fish oils. For example,distillation has been tried to reduce cholesterol from fish oils butsuch methods usually require extreme temperature and pressure. Moreover,distillation can produce dark oil and can adversely affect the taste ofthe oil. Also, distillation can be costly on a commercial scale, andyields are often poor. Other methods involving chemical extractionsand/or transformations have also been tried but the expense of reagentsand contamination from the chemical extractants, solvents, and reactantscan be problematic.

In light of the problems associated with removing undesirable compoundsfrom fish oils, what is needed in the art are methods for removingand/or reducing various compounds from oils. Also needed are oilsprepared from such methods (e.g., oils with reduced levels of variousundesirable compounds). The subject matter disclosed) herein meets theseand other needs.

SUMMARY

In accordance with the purposes of the disclosed materials, compounds,compositions, articles, and methods, as embodied and broadly describedherein, the disclosed subject matter, in one aspect, relates tocompositions and methods for preparing and using such compositions. In afurther aspect, the disclosed subject matter relates to methods ofremoving compounds (e.g., sterols) from compositions such as marineoils. In still a further aspect, the disclosed subject matter relates tomethods of bleaching compositions. Still further, the disclosed subjectmatter relates to compositions prepared by the methods disclosed herein.

Additional advantages will be set forth in part in the description thatfollows, and in part will be obvious from the description, or may belearned by practice of the aspects described below. The advantagesdescribed below will be realized and attained by means of the elementsand combinations particularly pointed out in the appended claims. It isto be understood that both the foregoing general description and thefollowing detailed description are exemplary and explanatory only andare not restrictive.

DETAILED DESCRIPTION

The materials, compounds, compositions, articles, and methods describedherein may be understood more readily by reference to the followingdetailed description of specific aspects of the disclosed subject matterand the Examples included therein.

Before the present materials, compounds, compositions, articles, andmethods are disclosed and described, it is to be understood that theaspects described below are not limited to specific synthetic methods orspecific reagents, as such may, of course, vary. It is also to beunderstood that the terminology used herein is for the purpose ofdescribing particular aspects only and is not intended to be limiting.

Also, throughout this specification, various publications arereferenced. The disclosures of these publications in their entiretiesare hereby incorporated by reference into this application in order tomore fully describe the state of the art to which the disclosed matterpertains. The references disclosed are also individually andspecifically incorporated by reference herein for the material containedin them that is discussed in the sentence in which the reference isrelied upon.

GENERAL DEFINITIONS

In this specification and in the claims that follow, reference will bemade to a number of terms, which shall be defined to have the followingmeanings:

Throughout the description and claims of this specification the word“comprise” and other forms of the word, such as “comprising” and“comprises,” means including but not limited to, and is not intended toexclude, for example, other additives, components, integers, or steps.

As used in the description and the appended claims, the singular forms“a,” “an,” and “the” include plural referents unless the context clearlydictates otherwise. Thus, for example, reference to “a compound”includes mixtures of two or more such compounds, reference to “anadsorbent” includes mixtures of two or more such adsorbents, referenceto “the oil” includes mixtures of two or more such oils, and the like.

“Optional” or “optionally” means that the subsequently described eventor circumstance can or cannot occur, and that the description includesinstances where the event or circumstance occurs and instances where itdoes not.

Ranges can be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, another aspect includes from the one particular value and/orto the other particular value. Similarly, when values are expressed asapproximations, by use of the antecedent “about,” it will be understoodthat the particular value forms another aspect. It will be furtherunderstood that the endpoints of each of the ranges are significant bothin relation to the other endpoint, and independently of the otherendpoint. It is also understood that there are a number of valuesdisclosed herein, and that each value is also herein disclosed as“about” that particular value in addition to the value itself. Forexample, if the value “10” is disclosed, then “about 10” is alsodisclosed. It is also understood that when a value is disclosed that“less than or equal to” the value, “greater than or equal to the value,”and possible ranges between values are also disclosed, as appropriatelyunderstood by the skilled artisan. For example, if the value “10” isdisclosed, then “less than or equal to 10” as well as “greater than orequal to 10” is also disclosed. It is also understood that throughoutthe application data is provided in a number of different formats andthat this data represents endpoints and starting points and ranges forany combination of the data points. For example, if a particular datapoint “10” and a particular data point “15” are disclosed, it isunderstood that greater than, greater than or equal to, less than, lessthan or equal to, and equal to 10 and 15 are considered disclosed aswell as between 10 and 15. It is also understood that each unit betweentwo particular units are also disclosed. For example, if 10 and 15 aredisclosed, then 11, 12, 13, and 14 are also disclosed.

References in the specification and concluding claims to parts by weightof a particular component in a composition denotes the weightrelationship between the component and any other components in thecomposition for which a part by weight is expressed. Thus, in a compoundcontaining 2 parts by weight of component X and 5 parts by weightcomponent Y, X and Y are present at a weight ratio of 2:5, and arepresent in such ratio regardless of whether additional components arecontained in the compound.

A weight percent (wt. %) of a component, unless specifically stated tothe contrary, is based on the total weight of the formulation orcomposition in which the component is included.

Reference will now be made in detail to specific aspects of thedisclosed materials, compounds, compositions, articles, and methods,examples of which are illustrated in the accompanying Examples.

Materials and Compositions

Disclosed herein are materials, compounds, compositions, and componentsthat can be used for, can be used in conjunction with, can be used inpreparation for, or are products of the disclosed methods andcompositions. These and other materials are disclosed herein, and it isunderstood that when combinations, subsets, interactions, groups, etc.of these materials are disclosed that while specific reference of eachvarious individual and collective combinations and permutation of thesecompounds may not be explicitly disclosed, each is specificallycontemplated and described herein. For example, if a compound isdisclosed and a number of modifications that can be made to a number ofcomponents or residues of the compound are discussed, each and everycombination and permutation that are possible are specificallycontemplated unless specifically indicated to the contrary. Thus, if aclass of components A, B, and C are disclosed as well as a class ofcomponents D, E, and F and an example of a combination composition A-Dis disclosed, then even if each is not individually recited, each isindividually and collectively contemplated. Thus, in this example, eachof the combinations A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F arespecifically contemplated and should be considered disclosed fromdisclosure of A, B, and C; D, E, and F; and the example combination A-D.Likewise, any subset or combination of these is also specificallycontemplated and disclosed. Thus, for example, the sub-group of A-E,B-F, and C-E are specifically contemplated and should be considereddisclosed from disclosure of A, B, and C; D, B, and F; and the examplecombination A-D. This concept applies to all aspects of this disclosureincluding, but not limited to, steps in methods of making and using thedisclosed compositions. Thus, if there are a variety of additional stepsthat can be performed it is understood that each of these additionalsteps can be performed with any specific aspect or combination ofaspects of the disclosed methods, and that each such combination isspecifically contemplated and should be considered disclosed.

Disclosed herein, in one aspect, are compositions that are oils. In manyexamples, the compositions are derived from marine oils. Marine oils, asused herein, refer to oils that are isolated from marine life. Forexample, marine oils can be oils that are isolated from fish, Molluscasuch as squid, cuttle fish, and/or octopus, Crustacea such as krill, andmarine mammals such as seals and whales. Specific examples of suitablemarine oils include, but are not limited to, Atlantic fish oils, Pacificfish oils, Mediterranean fish oils, light pressed fish oil, alkalinetreated fish oil, heat treated fish oil, light and heavy brown fish oil,tuna oil, bonito oil, sea bass oil, halibut oil, spearfish oil,barracuda oil, cod oil, menhaden oil, sardine oil, pilchard oil, anchovyoil, capelin oil, Atlantic cod oil, Atlantic herring oil, Atlanticmackerel oil, Atlantic menhaden oil, salmonids oil, shark oil, squidoil, octopus oil, krill oil, seal oil, whale oil, and the like,including mixtures and combinations thereof. Any marine oil andcombination of marine oil can be used in the disclosed compositions andmethods. In other examples, the compositions can be oils that areisolated from vegetables and plants, animals, and microbes, as well asedible oils. Further examples of suitable oils include crude oils fromsuch sources disclosed herein as well as semi-refined (also calledalkaline refined) and refined oils from such sources. Still further, thedisclosed compositions and methods can use oils comprising re-esterifiedtriglycerides. Also, any combination of these oils can be used.

The disclosed compositions can be prepared by methods described herein.For example, the disclosed methods can be used to reduce and/or removevarious compounds from oils to provide the disclosed compositions. Asone specific example, the disclosed compositions can be marine oils thathave been treated according to the disclosed methods and comprisereduced levels of compounds such as sterols, as compared to the originaloil. In other examples, the disclosed compositions can be plant,animals, microbe, edible, crude, semi-refined, refined, and/orre-esterified oils that have been treated according to the disclosedmethods, which comprise reduced levels of compounds such as sterols ascompared to the starting oil.

Levels of Sterols

In many examples, the disclosed compositions contain reduced levels ofsterols (e.g., cholesterol) as compared to a starting oil. By startingoil is meant any oil before it has been treated according to the methodsdisclosed herein. A suitable starting oil can be any oil as disclosedherein, for example, oils isolated from marine life, plant life, animallife, microbes, edible oils, as well as crude oils and such oils thathave been semi-refined, refined, and/or re-esterified.

The sterols that can be present at reduced levels in the disclosedcompositions or that are removed from the disclosed compositions, ascompared to a starting oil, can be cholesterols (e.g., free cholesteroland esterified cholesterol) and phytosterols.

In specific examples, the disclosed compositions can comprise less thanor equal to about 6.0 milligrams of sterol per gram of the composition.Also, disclosed are compositions that can comprise less than or equal toabout 5.9, 5.8, 5.7, 5.6, 5.5, 5.4, 5.3, 5.2, 5.1, 5.0, 4.9, 4.8, 4.7,4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3,3.2, 3.1, 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9,1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, 0.5,0.4, 0.3, 0.2, 0.1, or 0.0 milligrams of sterol per gram of thecomposition, where any of the stated values can form an upper and/orlower endpoint as appropriate. In other examples, the disclosedcompositions can comprise less than about 2.5, more specifically lessthan about 2.0 milligrams of sterol per gram of the composition.

The disclosed compositions, in further examples, can comprise from about6.0 to about 1.0, from about 6.0 to about 0.8, from about 6.0 to about0.6, from about 6.0 to about 0.4, from about 6.0 to about 0.2, fromabout 6.0 to about 0.0, from about 5.8 to about 1.0, from about 5.8 toabout 0.8, from about 5.8 to about 0.6, from about 5.8 to about 0.4,from about 5.8 to about 0.2, from about 5.8 to about 0.0, from about 5.6to about 1.0, from about 5.6 to about 0.8, from about 5.6 to about 0.6,from about 5.6 to about 0.4, from about 5.6 to about 0.2, from about 5.6to about 0.0, from about 5.4 to about 1.0, from about 5.4 to about 0.8,from about 5.4 to about 0.6, from about 5.4 to about 0.4, from about 5.4to about 0.2, from about 5.4 to about 0.0, from about 5.2 to about 1.0,from about 5.2 to about 0.8, from about 5.2 to about 0.6, from about 5.2to about 0.4, from about 52 to about 0.2, from about 5.2 to about 0.0,from about 5.0 to about 1.0, from about 5.0 to about 0.8, from about 5.0to about 0.6, from about 5.0 to about 0.4, from about 5.0 to about 0.2,from about 5.0 to about 0.0, from about 4.8 to about 1.0, from about 4.8to about 0.8, from about 4.8 to about 0.6, from about 4.8 to about 0.4,from about 4.8 to about 0.2, from about 4.8 to about 0.0, from about 4.6to about 1.0, from about 4.6 to about 0.8, from about 4.6 to about 0.6,from about 4.6 to about 0.4, from about 4.6 to about 0.2, from about 4.6to about 0.0, from about 4.4 to about 1.0, from about 4.4 to about 0.8,from about 4.4 to about 0.6, from about 4.4 to about 0.4, from about 4.4to about 0.2, from about 4.4 to about 0.0, from about 4.2 to about 1.0,from about 4.2 to about 0.8, from about 4.2 to about 0.6, from about 4.2to about 0.4, from about 4.2 to about 0.2, from about 4.2 to about 0.0,from about 4.0 to about 1.0, from about 4.0 to about 0.8, from about 4.0to about 0.6, from about 4.0 to about 0.4, from about 4.0 to about 0.2,from about 4.0 to about 0.0, from about 3.8 to about 1.0, from about 3.8to about 0.8, from about 3.8 to about 0.6, from about 3.8 to about 0.4,from about 3.8 to about 0.2, from about 3.8 to about 0.0, from about 3.6to about 1.0, from about 3.6 to about 0.8, from about 3.6 to about 0.6,from about 3.6 to about 0.4, from about 3.6 to about 0.2, from about 3.6to about 0.0, from about 3.4 to about 1.0, from about 3.4 to about 0.8,from about 3.4 to about 0.6, from about 3.4 to about 0.4, from about 3.4to about 0.2, from about 3.4 to about 0.0, from about 3.2 to about 1.0,from about 3.2 to about 0.8, from about 3.2 to about 0.6, from about 3.2to about 0.4, from about 3.2 to about 0.2, from about 3.2 to about 0.0,from about 3.0 to about 1.0, from about 3.0 to about 0.8, from about 3.0to about 0.6, from about 3.0 to about 0.4, from about 3.0 to about 0.2,from about 3.0 to about 0.0, from about 2.8 to about 1.0, from about 2.8to about 0.8, from about 2.8 to about 0.6, from about 2.8 to about 0.4,from about 2.8 to about 0.2, from about 12.8 to about 0.0, from about2.6 to about 1.0, from about 2.6 to about 0.8, from about 2.6 to about0.6, from about 2.6 to about 0.4, from about 2.6 to about 0.2, fromabout 2.6 to about 0.0, from about 2.4 to about 1.0, from about 2.4 toabout 0.8, from about 2.4 to about 0.6, from about 2.4 to about 0.4,from about 2.4 to about 0.2, from about 2.4 to about 0.0, from about 2.2to about 1.0, from about 2.2 to about 0.8, from about 2.2 to about 0.6,from about 2.2 to about 0.4, from about 2.2 to about 0.2, from about 2.2to about 0.0, from about 2.0 to about 1.0, from about 2.0 to about 0.8,from about 2.0 to about 0.6, from about 2.0 to about 0.4, from about 2.0to about 0.2, from about 2.0 to about 0.0, from about 1.8 to about 1.0,from about 1.8 to about 0.8, from about 1.8 to about 0.6, from about 1.8to about 0.4, from about 1.8 to about 0.2, from about 1.8 to about 0.0,from about 1.6 to about 1.0, from about 1.6 to about 0.8, from about 1.6to about 0.6, from about 1.6 to about 0.4, from about 1.6 to about 0.2,from about 1.6 to about 0.0, from about 1.4 to about 1.0, from about 1.4to about 0.8, from about 1.4 to about 0.6, from about 1.4 to about 0.4,from about 1.4 to about 0.2, from about 1.4 to about 0.0, from about 1.2to about 1.0, from about 1.2 to about 0.8, from about 1.2 to about 0.6,from about 1.2 to about 0.4, from about 1.2 to about 0.2, from about 1.2to about 0.0, from about 1.0 to about 0.8, from about 1.0 to about 0.6,from about 1.0 to about 0.4, from about 1.0 to about 0.2, or from about1.0 to about 0.0 milligrams of sterol per gram of the composition.

In still other examples, the disclosed compositions can comprise fromabout 6.0 to about 2.0, from about 6.0 to about 1.7, from about 6.0 toabout 1.5, from about 6.0 to about 1.3, from about 5.8 to about 2.0,from about 5.8 to about 1.7, from about 5.8 to about 1.5, from about 5.8to about 1.3, from about 5.6 to about 2.0, from about 5.6 to about 1.7,from about 5.6 to about 1.5, from about 5.6 to about 1.3, from about 5.4to about 2.0, from about 5.4 to about 1.7, from about 5.4 to about 1.5,from about 5.4 to about 1.3, from about 5.2 to about 2.0, from about 5.2to about 1.7, from about 5.2 to about 1.5, from about 5.2 to about 1.3,from about 5.0 to about 2.0, from about 5.0 to about 1.7, from about 5.0to about 1.5, from about 5.0 to about 13, from about 4.8 to about 2.0,from about 4.8 to about 1.7, from about 4.8 to about 1.5, from about 4.8to about 1.3, from about 4.6 to about 2.0, from about 4.6 to about 1.7,from about 4.6 to about 1.5, from about 4.6 to about 1.3, from about 4.4to about 2.0, from about 4.4 to about 1.7, from about 4.4 to about 1.5,from about 4.4 to about 1.3, from about 4.2 to about 2.0, from about 4.2to about 1.7, from about 4.2 to about 1.5, from about 4.2 to about 1.3,from about 4.0 to about 2.0, from about 4.0 to about 1.7, from about 4.0to about 1.5, from about 4.0 to about 1.3, from about 3.8 to about 2.0,from about 3.8 to about 1.7, from about 3.8 to about 1.5, from about 3.8to about 1.3, from about 3.6 to about 2.0, from about 3.6 to about 1.7,from about 3.6 to about 1.5, from about 3.6 to about 1.3, from about 3.4to about 2.0, from about 3.4 to about 1.7, from about 3.4 to about 1.5,from about 3.4 to about 1.3, from about 3.2 to about 2.0, from about 3.2to about 1.7, from about 3.2 to about 1.5, from about 3.2 to about 1.3,from about 3.0 to about 2.0, from about 3.0 to about 1.7, from about 3.0to about 1.5, from about 3.0 to about 1.3, from about 2.8 to about 2.0,from about 2.8 to about 1.7, from about 2.8 to about 1.5, from about 2.8to about 1.3, from about 2.6 to about 2.0, from about 2.6 to about 1.7,from about 2.6 to about 1.5, from about 2.6 to about 1.3, from about 2.4to about 2.0, from about 2.4 to about 1.7, from about 2.4 to about 1.5,from about 2.4 to about 1.3, from about 2.2 to about 2.0, from about 2.2to about 1.7, from about 2.2 to about 1.5, from about 2.2 to about 1.3,from about 2.0 to about 1.7, from about 2.0 to about 1.5, or from about2.0 to about 1.3 milligrams of sterol per gram of the composition.

In yet other examples, the disclosed compositions can comprise fromabout 6.0 to about 3.0, from about 6.0 to about 2.8, from about 6.0 toabout 2.6, from about 6.0 to about 2.4, from about 6.0 to about 2.2,from about 5.8 to about 3.0, from about 5.8 to about 2.8, from about 5.8to about 2.6, from about 5.8 to about 2.4, from about 5.8 to about 2.2,from about 5.6 to about 3.0, from about 5.6 to about 2.8, from about 5.6to about 2.6, from about 5.6 to about 2.4, from about 5.6 to about 2.2,from about 5.4 to about 3.0, from about 5.4 to about 2.8, from about 5.4to about 2.6, from about 5.4 to about 2.4, from about 5.4 to about 2.2,from about 5.2 to about 3.0, from about 5.2 to about 2.8, from about 5.2to about 2.6, from about 5.2 to about 2.4, from about 5.2 to about 2.2,from about 5.0 to about 3.0, from about 5.0 to about 2.8, from about 5.0to about 2.6, from about 5.0 to about 2.4, from about 5.0 to about 2.2,from about 4.8 to about 3.0, from about 4.8 to about 2.8, from about 4.8to about 2.6, from about 4.8 to about 2.4, from about 4.8 to about 2.2,from about 4.6 to about 3.0, from about 4.6 to about 2.8, from about 4.6to about 2.6, from about 4.6 to about 2.4, from about 4.6 to about 2.2,from about 4.4 to about 3.0, from about 4.4 to about 2.8, from about 4.4to about 2.6, from about 4.4 to about 2.4, from about 4.4 to about 2.2,from about 4.2 to about 3.0, from about 4.2 to about 2.8, from about 4.2to about 2.6, from about 4.2 to about 2.4, from about 4.2 to about 2.2,from about 4.0 to about 3.0, from about 4.0 to about 2.8, from about 4.0to about 2.6, from about 4.0 to about 2.4, from about 4.0 to about 2.2,from about 3.8 to about 3.0, from about 3.8 to about 2.8, from about 3.8to about 2.6, from about 3.8 to about 2.4, from about 3.8 to about 2.2,from about 3.6 to about 3.0, from about 3.6 to about 2.8, from about 3.6to about 2.6, from about 3.6 to about 2.4, from about 3.6 to about 2.2,from about 3.4 to about 3.0, from about 3.4 to about 2.8, from about 3.4to about 2.6, from about 3.4 to about 2.4, from about 3.4 to about 2.2,from about 3.2 to about 3.0, from about 3.2 to about 2.8, from about 3.2to about 2.6, from about 3.2 to about 2.4, from about 3.2 to about 2.2,from about 3.0 to about 2.8, from about 3.0 to about 2.6, from about 3.0to about 2.4, of from about 3.0 to about 2.2 milligrams of sterol pergram of the composition.

In a particular example, the amount of sterol in the disclosedcomposition can be from about 1.2 to about 0.8, from about 1.1 to about0.9, or about 1.0 milligrams per gram of the composition.

Color

The disclosed compositions can also be lighter in color that crude oil,even crude oil that has been subjected to a bleaching procedure. Colorsfor the disclosed compositions and crude oils can be defined by theirGardner color. Gardner color is a color measurement based on thechromaticities of glass standards numbered from 1 for the lightest to 18for the darkest. Gardner color measurements are well known and caninvolve an apparatus known as a comparator, such as the Comparator 3000from Tintometer Ltd. (United Kingdom). Methods for measuring the Gardnercolor, of the disclosed compositions are disclosed in ASTM D 1544; thistoothed is available from ASTM. International and is incorporated byreference herein in its entirety.

In some specific examples, the disclosed compositions can have a Gardnercolor of less than or equal to about 18. In other examples, thecompositions disclosed herein can have a Gardner color of less than orequal to about 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3,2, or 1 as determined by ASTM D 1544, where any of the stated values canform an upper and/or lower endpoint as appropriate.

In other examples of the disclosed compositions, the Gardner color canbe from about 18 to about 1, from about 17 to about 1, from about 16 toabout 1, from about 15 to about 1, from about 14 to about 1, from about13 to about 1, from about 12 to about 1, from about 11 to about 1, fromabout 10 to about 1, from about 9 to about 1, from about 8 to about 1,from about 7 to about 1, from about 6 to about 1, from about 5 to about1, from about 4 to about 1, from about 3 to about 1, from about 2 toabout 1, from about 18 to about 2, from about 17 to about 2, from about16 to about 2, from about 15 to about 2, from about 14 to about 2, fromabout 13 to about 2, from about 12 to about 2, from about 11 to about 2,from about 10 to about 2, from about 9 to about 2, from about 8 to about2, from about 7 to about 2, from about 6 to about 2, from about 5 toabout 2, from about 4 to about 2, from about 3 to about 2, from about 18to about 3, from about 17 to about 3, from about 16 to about 3, fromabout 15 to about 3, from about 14 to about 3, from about 13 to about 3,from about 12 to about 3, from about 11 to about 3, from about 10 toabout 3, from about 9 to about 3, from about 8 to about 3, from about 7to about 3, from about 6 to about 3, from about 5 to about 3, from about4 to about 3, from about 18 to about 4, from about 17 to about 4, fromabout 16 to about 4, from about 15 to about 4, from about 14 to about 4,from about 13 to about 4, from about 12 to about 4, from about 11 toabout 4, from about 10 to about 4, from about 9 to about 4, from about 8to about 4, from about 7 to about 4, from about 6 to about 4, from about5 to about 4, from about 18 to about 5, from about 17 to about 5, fromabout 16 to about 5, from about 15 to about 5, from about 14 to about 5,from about 13 to about 5, from about 12 to about 5, from about 11 toabout 5, from about 10 to about 5, from about 9 to about 5, from about 8to about 5, from about 7 to about 5, from about 6 to about 5, from about18 to about 6, from about 17 to about 6, from about 16 to about 6, fromabout 15 to about 6, from about 14 to about 6, from about 13 to about 6,from about 12 to about 6, from about 11 to about 6, from about 10 toabout 6, from about 9 to about 6, from about 8 to about 6, from about 7,to about 6, from about 18 to about 7, from about 17 to about 7, fromabout 16 to about 7, from about 15 to about 7, from about 14 to about 7,from about 13 to about 7, from about 12 to about 7, from about 11 toabout 7, from about 10 to about 7, from about 9 to about 7, from about 8to about 7, from about 18 to about 8, from about 17 to about 8, fromabout 16 to about 8, from about 15 to about 8, from about 14 to but 8,from about 13 to about 8, from about 12 to about 8, from about 11 toabout 8, from about 10 to about 8, from about 9 to about 8, from about18 to about 9, from about 17 to about 9, from about 16 to about 9, fromabout 15 to about 9, from about 14 to about 9, from about 13 to about 9,from about 12 to about 9, from about 11 to about 9, from about 10 toabout 9, from about 18 to about 10, from about 17 to about 10, fromabout 16 to about 10, from about 1.5 to about 10, from about 14 to about10, from about 13 to about 10, from about 12 to about 10, from about 11to about 10, from about 18 to about 11, from about 17 to about 11, fromabout 16 to about 11, from about 15 to about 11, from about 14 to about11, from about 13 to about 11, from about 12 to about 11, from about 18to about 12, from about 17 to about 12, from about 16 to about 12, fromabout 15 to about 12, from about 14 to about 12, from about 13 to about12, from about 18 to about 13, from about 17 to about 13, from about 16to about 13, from about 15 to about 13, from about 14 to about 13, fromabout 18 to about 14, from about 17 to about 14, from about 16 to about14, from about 15 to about 14, from about 18 to about 15, from about 17to about 15, from about 16 to about 15, from about 18 to about 16, fromabout 17 to about 16, or from about 18 to about 17 as determined by ASTMD 1544.

Fatty Acids

The disclosed compositions can contain one or more fatty acids. By“fatty acid” is meant a carboxylic acid with at least 8 carbon atoms.“Fatty acid” is also meant to include a residue of a fatty acid. Theterm “residue” as used herein refers to the moiety that is the resultingproduct of the specified chemical species in a particular reactionscheme or subsequent formulation or chemical product, regardless ofwhether the moiety is actually obtained from the specified chemicalspecies. For example, an “unsaturated fatty acid residue” refers to themoiety which results when an unsaturated fatty acid participates in aparticular reaction (e.g., the residue can be an unsaturated fatty acylgroup RCO— or unsaturated acyloxyl group RCOO—, where R is anunsaturated chain). In this case, the unsaturated fatty acid residue is“derived” from the unsaturated fatty acid. It is understood that thismoiety can be obtained by a reaction with a species other than thespecified unsaturated fatty acid, for example, by a reaction with anunsaturated fatty acid chloride, ester, or anhydride.

In some examples, the fatty acids and residues thereof that can bepresent in the disclosed compositions can comprise at least 8, at least10, at least 12, at least 14, at least 16, at least 18, or at least 20carbon atoms. In some other examples, the fatty acids or residuesthereof can contain about 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,38, 39, 40, 41, 42, 43, 44, or 45 carbon atoms, where any of the statedvalues can form an upper and/or lower endpoint as appropriate. In stillother examples, the fatty acids and residues thereof can comprise amixture of fatty acids and residues thereof having a range of carbonatoms. For example, the fatty acids and residues thereof can comprisefrom about 8 to about 40, from about 10 to about 38, from about 12 toabout 36, from about 14 to about 34, from about 16 to about 32, fromabout 18 to about 30, or from about 20 to about 28 carbon atoms.

The fatty acids and residues thereof that can be present in thedisclosed compositions can be saturated, unsaturated, or a mixture ofsaturated and unsaturated fatty acids. By “saturated” is meant that themolecule or residue contains no carbon-carbon double or triple bounds.By “unsaturated” is meant that the molecule or residue contains at leastone carbon-carbon double or triple bond. The disclosed compositions canalso be processed to result in a particular mixture of fatty acids(e.g., having only saturated fatty acids, only unsaturated fatty acids,mixtures of both saturated and unsaturated fatty acids, or mixtures offatty acids of a certain chain length or range of chain lengths).

In many examples, the disclosed compositions are derived from marineoils, plant oils, or animal oils and contain various fatty acids andresidues thereof, examples of which are disclosed herein. As noted, anysuch oil can be used in the disclosed compositions and methods. It isalso contemplated that while a particular fatty acid may not be presentin the crude oil from which a specific composition is derived, such afatty acid can be added to the composition at any time (e.g., prior,during, or after the methods disclosed herein).

Unsaturated Fatty Acids

The unsaturated fatty acids and residues thereof that can be present inthe disclosed compositions contain at least one unsaturated bond (i.e.,a carbon-carbon double or triple bond). In one example, the unsaturatedfatty acids and residues thereof can comprise at least 2, at least 3, atleast 4, at least 5, at least 6, at least 7, or at least 8 carbon-carbondouble bonds, triple bonds, or any combination thereof. In anotherexample, the unsaturated fatty acids or residues thereof can comprise 1,2, 3, 4, 5, 6, 7, or 8 unsaturated bonds, where any of the stated valuescan form an upper and/or lower endpoint as appropriate.

Monoene Acids and Residues

In one aspect, the unsaturated fatty acids or residues thereof cancomprise one carbon-carbon double bond (i.e., a monoene acid orresidue). Examples of unsaturated fatty acids and residues thereof thatcan be present in the disclosed compositions include, but are notlimited to, those in the following Table 1.

TABLE 1 Examples of Monoenes Total number of carbon Carbon number wheredouble bond begins. atoms in the fatty acid or (“c” denotes a cis doublebond; “t” denotes residue chain. a trans double bond) 10 4c 12 4c 14 4cand 9c 16 3t, 4c, 5t, 6c, 6t, 9c (palmitooleic), and 11c 18 3t, 5c, 5t,6c (petroselinic), 6t, 9c (oleic), 10c, 11c (cis-vaccenic), 11t(vaccenic), and 13c 20 5c, 9c (gadolenic), 11c, 13c, and 15c 22 5c, 11c(cetoleic), 13c (erucic), and 15c 24 15c (selacholeic, nervonic) 26 9c,and 17c (ximenic) 28 9c, 19c (lumequic) 30 21c

Polyene Acids and Residues (Methylene Interrupted)

In other examples, the unsaturated fatty acids and residues thereof cancomprise at least two unsaturated bonds (e.g., polyene acids orresidues). In some examples, the unsaturated fatty acids and residuesthereof can comprise at least one pair of methylene interruptedunsaturated bonds. By “methylene interrupted unsaturated bond” is meantthat one carbon-carbon double or triple bond is separated from anothercarbon-carbon double or triple bond by at least one methylene group(i.e., CH₂). Specific examples of unsaturated fatty acids that containat least one pair of methylene interrupted unsaturated bonds include,but are not limited to, the n-1 family derived from 9, 12, 15-16:3; n-2family derived from 9, 12, 15-17:3, 15:3, 17:3, 17:4, 20:4; n-3 familyderived from 9, 12, 15-18:3, 15:2, 15:3, 15:4, 16:3, 16:4, 18:3(α-linolenic), 18:4, 18:5, 20:2, 20:3, 20:4, 20:5 (EPA), 21:5, 22:3,22:5 (DPA), 22:6 (DHA), 24:3, 24:4, 24:5, 24:6, 26:5, 26:6, 28:7, 30:5;n-4 family derived from 9, 12-16:2, 16:2, 16:3, 18:2, 18:3; n-5 familyderived from 9, 12-17:2, 15:2, 17:2, 17:3, 19:2, 19:4, 20:3, 20:4, 21:4,21:5; n-6 family derived from 9, 12-18:2, 15:2, 16:2, 18:2 (linoleicacid), 18:3 (γ-linolenic acid); 20:2, 20:3, 20:4 (arachidonic acid),22:2, 22:3, 22:4 (adrenic acid), 22:5, 24:2, 24:4, 25:2, 26:2, 30:4; n-7family derived front 9-16:1, 15:2, 16:2, 17:2, 18:2, 19:2; n-8 familyderived from 9-17:1, 15:2, 16:2, 17:2, 18:2, 19:2; n-9 family derivedfrom 9-18:1, 17:2, 18:2, 20:2, 20:3, 22:3, 22:4; n-11 family 19:2, andthe n-12 family 20:2.

In the above paragraph (and throughout) the compounds are identified byreferring first to the “n-x family,” where x is the position in thefatty acid where the first double bond begins. The numbering schemebegins at the terminal end of the fatty acid, where, for example, theterminal CH₃ group is designated position 1. In this sense, the n-3family would be an omega-3 fatty acid, as described herein. The nextnumber identifies the total number of carbon atoms in the fatty acid.The third number, which is after the colon, designates the total numberof double bonds in the fatty acid. So, for example, in the n-1 family,16:3, refers to a 16 carbon long fatty acid with 3 double bonds, eachseparated by a methylene, wherein the first double bond begins atposition 1, i.e., the terminal end of the fatty acid. In anotherexample, in the n-6 family, 18:3, refers to an 18 carbon long fatty acidwith 3 methylene separated double bonds beginning at position 6, i.e.,the sixth carbon from the terminal end of the fatty acid, and so forth.

Some other examples are fatty acids and residues thereof that contain atleast one pair of unsaturated bonds interrupted by more than onemethylene group. Suitable examples of these acids and residues thereofinclude, but are not limited to, those in the following Table 2.

TABLE 2 Examples of Polyene Acids and Residues with Double BondsInterrupted by Several Methylene Units Total number of carbon Carbonnumber where double bond begins. atoms in the fatty acid or (“c” denotesa cis double bond; “t” denotes a residue chain. trans double bond) 18 5,9 5, 11 2t, 9, 12 3t, 9, 12 5t, 9, 12 5, 9, 12 5, 11, 14 3t, 9, 12, 155, 9, 12, 15 20 5, 11 5, 13 7, 11 7, 13 5, 11, 14 7, 11, 14 5, 11, 14,17 22 5, 11 5, 13 7, 13 7, 15 7, 17 9, 13 9, 15

Polyene Acid and Residues (Conjugated)

Still other examples of unsaturated fatty acids and residues thereofthat can be present in the disclosed compositions are those that containat least one conjugated unsaturated bond. By “conjugated unsaturatedbond” is meant that at least one pair of carbon-carbon double and/ortriple bonds are bonded together, without a methylene (CH₂) groupbetween them (e.g., —CH═CH—CH═CH—). Specific examples of unsaturatedfatty acids that contain conjugated unsaturated bonds include, but arenot limited to, those in the following Table 3.

TABLE 3 Examples of Conjugated Polyene Acids and Residues Total numberof carbon Carbon number where double bond begins. atoms in the fattyacid or (“c” denotes a cis double bond; “t” denotes a residue chain.trans double bond) 10 2t, 4t, 6c 2c, 4t, 6t 3t, 5t, 7c 3c, 5t, 7t 12 3,5, 7, 9, 11 14 3, 5, 7, 9, 11 18 10t, 12t 8c, 10t, 12c (jacaric) 8t,10t, 12c (calendic) 8t, 10t, 12t 9t, 11t, 13c (catalpic) 9c, 11t, 13t(α-eleostearic) 9c, 11t, 13c (punieic) 9t, 11t, 13t (β-eleostearic) 9c,11t, 13t, 15c (α-parinaric) 9t, 11t, 13t, 15t (β-parinaric)

Omega-3 Fatty Acids

In many examples, the disclosed compositions can comprise one or moreomega-3 fatty acids or a residue thereof. An omega-3 fatty acid is anunsaturated fatty acid that contains as its terminus CH₂—CH₂—CH═CH—.Specific examples of omega-3 fatty acids that can be present in thedisclosed compositions include, but are not limited to, linolenic acid(18:3ω3), octadecatetraenoic acid (18:4ω3), eicosapentaenoic acid(20:5ω3) (EPA), docosahexaenoic acid (22:6ω3) (MIA), docosapentaenoicacid (22:6ω3) (DPA), including residues, derivatives, and mixturesthereof.

In still other examples, the unsaturated fatty acids or residues thereofcan be derived from a compound comprising the following formula:

wherein R¹ is a C₃-C₄₀ alkyl or alkenyl group comprising at least onedouble bond. The term “alkane” or “alkyl” as used herein is a saturatedhydrocarbon group. The term “alkene” or “alkenyl” as used herein is ahydrocarbon group containing at least one carbon-carbon double bond.Asymmetric structures such as (AB)C═C(CD) are intended to include boththe E and Z isomers (cis and trans). This may be presumed in structuralformulae herein wherein an asymmetric alkene is present, or it flay beexplicitly indicated by the bond symbol C═C. In a further example, R¹can be a C₅-C₃₈, C₆-C₃₆, C₈-C₃₄, C₁₀-C₃₂, C₁₂-C₃₀, C₁₄-C₂₈, or C₁₆-C₂₆,C₁₈-C₂₄ alkenyl group. In yet another example, the alkenyl group of R¹can have from 2 to 6, from 3 to 6, from 4 to 6, or from 5 to 6 doublebonds. Still further, the alkenyl group of R¹ can have from 1, 2, 3, 4,5, or 6 double bonds, where any of the stated values can form an upperor lower endpoint when appropriate.

Exemplary Unsaturated Fatty Acid

Some specific examples of unsaturated fatty acids and residues derivedtherefrom that can be present in the disclosed compositions include, butare not limited to, linoleic acid, linolenic acid, γ-linolenic acid,arachidonic acid, mead acid, stearidonic acid, α-eleostearic acid,eleostearic acid, pinolenic acid, docosadicnic acid, docosatetraenoicacid, docosapentaenoic acid, docosahexaenoic acid, octadecadienoic acid,octadecatrienoic acid, eicosatetraenoic acid, eicosapentaenoic, or anycombination thereof. In one aspect, the unsaturated fatty acid residuecan be derived from cicosapentaenoic acid 20:5ω3 (EPA), docosahexaenoicacid 22:6ω3 (DHA), docosapentaenoic acid 22:5ω3 (DPA), and anycombination thereof.

Amounts of DHA/EPA

As noted many of the disclosed compositions can contain the omega-3fatty acids EPA and DHA or a residue thereof. Each of these unsaturatedfatty acids or residues can be present in the disclosed compositions inan amount of from about 0 to about 700 milligrams per gram of thecomposition. In other examples, DHA and/or EPA can each be present in anamount of about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130,140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270,280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410,420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550,560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, or700 milligrams per gram of the composition, where any of the statedvalues can form an upper and/or lower endpoint as appropriate.

In still other examples, DHA and/or EPA can each be present in thedisclosed compositions in an amount from about 50 to about 700, fromabout 100 to about 700, from about 150 to about 700, from about 200 toabout 700, from about 250 to about 700, from about 300 to about 700,from about 350 to about 700, from about 400 to about 700, from about 450to about 700, from about 500 to about 700, from but 550 to about 700,from about 600 to about 700, from about 650 to about 700, from about 0to about 650, from about 50 to about 650, from about 100 to about 650,from about 150 to about 650, from about 200 to about 650, from about 250to about 650, from about 300 to about 650, from about 350 to about 650,from about 400 to about 650, from about 450 to about 650, from about 500to about 650, from about 550 to about 650, from about 600 to about 650,from about 0 to about 600, from about 50 to about 600, from about 100 toabout 600, from about 150 to about 600, from about 200 to about 600,from about 250 to about 600, from about 300 to about 600, from about 350to about 600, from about 400 to about 600, from about 450 to about 600,from about 500 to about 600, from about 550 to about 600, from about 0to about 550, from about 50 to about 550, from about 100 to about 550,from about 150 to about 550, from about 200 to about 550, from about 250to about 550, from about 300 to about 550, from about 350 to about 550,from about 400 to about 550, from about 450 to about 550, from about 500to about 550, from about 0 to about 500, from about 50 to about 500,from about 100 to about 500, from about 150 to about 500, from about 200to about 500, from about 250 to about 500, from about 300 to about 500,from about 350 to about 500, from about 400 to about 500, from about 450to about 500, from about 0 to about 450, from about 50 to about 450,from about 100 to about 450, from about 150 to about 450, from about 200to about 450, from about 250 to about 450, from about 300 to about 450,from about 350 to about 450, from about 400 to about 450, from about 0to about 400, from about 50 to about 400, from about 100 to about 400,from about 150 to about 400, from about 200 to about 400, from about 250to about 400, from about 300 to about 400, from about 350 to about 400,from about 0 to about 350, from about 50 to about 350, from about 100 toabout 350, from about 150 to about 350, from about 200 to about 350,from about 250 to about 350, from about 300 to about 350, from about 0to about 300, from about 50 to about 300, from about 100 to about 300,from about 150 to about 300, from about 200 to about 300, from about 250to about 300, from about 0 to about 250, from about 50 to about 250,from about 100 to about 250, from about 150 to about 250, from about 200to about 250, from about 0 to about 200, from about 50 to about 200,from about 100 to about 200, from about 150 to about 200, from about 0to about 150, from about 50 to about 150, from about 100 to about 150,from about 0 to about 100, from about 50 to about 100, from about 0 toabout 50 milligrams per gram of composition.

The amount of EPA and DHA that can be present in the disclosedcompositions can also be described in terms of weight % (wt. %). Forexample, the disclosed compositions can comprise from about 0 to about70 wt. % EPA and/or DHA, based on the total weight of the composition.In other examples, the disclosed compositions can comprise about 0, 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, or 70 wt. % based on thetotal weight of the composition, where any of the stated values can forman upper and/or lower endpoint as appropriate.

In still further examples, the amount of EPA and/or DHA that can bepresent in the disclosed composition can be from about 5 to about 70,from about 10 to about 70, from about 15 to about 70, from about 20 toabout 70, from about 25 to about 70, from about 30 to about 70, fromabout 35 to about 70, from about 40 to about 70, from about 45 to about70, from about 50 to about 70, from about 55 to about 70, from about 60to about 70, from about 65 to about 70, from about 0 to about 65, fromabout 5 to about 65, from about 10 to about 65, from about 15 to about65, from about 20 to about 65, from about 25 to about 65, from about 30to about 65, from about 35 to about 65, from about 40 to about 65, fromabout 45 to about 65, from about 50 to about 65, from about 55 to about65, from about 60 to about 65, from about 0 to about 60, from about 5 toabout 60, from about 10 to about 60, from about 15 to about 60, fromabout 20 to about 60, from about 25 to about 60, from about 30 to about60, from about 35 to about 60, from about 40 to about 60, from about 45to about 60, from about 50 to about 60, from about 55 to about 60, fromabout 0 to about 55, from about 5 to about 55, from about 10 to about55, from about 15 to about 55, from about 20 to about 55, from about 25to about 55, from about 30 to about 55, from about 35 to about 55, fromabout 40 to about 55, from about 45 to about 55, from about 50 to about55, from about 0 to about 50, from about 5 to about 50, from about 10 toabout 50, from about 15 to about 50, from about 20 to about 50, fromabout 25 to about 50, from about 30 to about 50, from about 35 to about50, from about 40 to about 50, from about 45 to about 50, from about 0to about 45, from about 5 to about 45, from about 10 to about 45, fromabout 15 to about 45, from about 20 to about 45, from about 25 to about45, from about 30 to about 45, from about 35 to about 45, from about 40to about 45, from about 0 to about 40, from about 5 to about 40, fromabout 10 to about 40, from about 1.5 to about 40, from about 20 to about40, from about 25 to about 40, from about 30 to about 40, from about 35to about 40, from about 0 to about 35, from about 5 to about 35, fromabout 10 to about 35, from about 15 to about 35, from about 20 to about35, from about 25 to about 35, from about 30 to about 35, from about 0to about 30, from about 5 to about 30, from about 10 to about 30, fromabout 15 to about 30, from about 20 to about 30, from about 25 to about30, from about 0 to about 25, from about 5 to about 25, from about 10 toabout 25, from about 15 to about 25, from about 20 to about 25, fromabout 0 to about 20, from about 5 to about 20, from about 10 to about20, from about 15 to about 20, from about 0 to about 15, from about 5 toabout 15, from about 10 to about 15, from about 0 to about 10, fromabout 5 to about 10, from about 0 to about 5 wt. % based on the totalweight of the composition. In some other specific examples, the amountof EPA and/or DHA that can be present in the disclosed compositions canbe about 0.3, 5, 12, 18, 25, or 60 wt. % based on the total weight ofthe composition, where any of the stated values can form an upper and/orlower endpoint when appropriate.

The amount of EPA and DHA present in the disclosed compositions can alsobe described in terms of the wt. % ratio of EPA to DHA. For example, thewt. % ratio of EPA to DHA in the disclosed compositions can be about18:12 (i.e., about 18 wt. % EPA to about 12 wt. % DHA, based on thetotal weight of the composition). Other wt. % ratios of EPA to DHA thatcan be present in the disclosed compositions include, but are notlimited to, about 5:25, about 60:0.3, and about 0.8:60. Further wt. %ratios of EPA to DHA for the disclosed compositions can be about 0:70,5:70, 10:70, 15:70, 20:70, 25:70, 30:70, 70:30, 70:25, 70:20, 70:15,70:10, 70:5, 70:0, 0:65, 5:65, 10:65, 15:65, 20:65, 25:65, 30:65, 35:65,65:35, 65:30, 65:25, 65:20, 65:15, 65:10, 65:5, 65:0, 0:60, 5:60, 10:60,15:60, 20:60, 25:60, 30:60, 35:60, 40:60, 60:40, 60:35, 60:30, 60:25,60:20, 60:15, 60:10, 60:5, 60:0, 0:55, 5:55, 10:55, 15:55, 20:55, 25:55,30:55, 35:55, 40:55, 45:55, 55:45, 55:40, 55:35, 55:30, 55:25, 55:20,55:15, 55:10, 55:5, 55:0, 0:50, 5:50, 10:50, 15:50, 20:50, 25:50, 30:50,35:50, 40:50, 45:50, 50:50, 50:45, 50:40, 50:35, 50:30, 50:25, 50:20,50:15, 50:10, 50:5, 50:0, 0:45, 5:45, 10:45, 15:45, 20:45, 25:45, 30:45,35:45, 40:45, 45:45, 45:40, 45:35, 45:30, 45:25, 45:20, 45:15, 45:10,45:5, 45:0, 0:40, 5:40, 10:40, 15:40, 20:40, 25:40, 30:40, 35:40, 40:40,40:35, 40:30, 40:25, 40:20, 40:15, 40:10, 40:5, 40:0, 0:35, 5:35, 10:35,15:35, 20:35, 25:35, 30:35, 35:35, 35:30, 35:25, 35:20, 35:15, 35:10,35:5, 35:0, 0:30, 5:30, 10:30, 15:30, 20:30, 25:30, 30:30, 30:25, 30:20,30:15, 30:10, 30:5, 30:0, 0:25, 5:25, 10:25, 15:25, 20:25, 25:25, 25:20,25:15, 25:10, 25:5, 25:0, 0:20, 5:20, 10:20, 15:20, 20:20, 20:15, 20:10,20:5, 20:0, 0:15, 5:15, 10:15, 15:15, 15:10, 15:5, 15:0, 0:10, 5:10,10:10, 10:5, 10:0, 0:5, 5:5, or 5:0.

Additional Fatty Acids

Examples of additional fatty acids and residues thereof that can bepresent in the disclosed compositions include, but are not limited to,the saturated fatty acids capric acid (C10), lauric acid (C12), myristicacid (C14), palmitic acid (C16), margaric acid (C17), stearic acid(C18), arachidic acid (C20), behenic acid (C22), lignoceric acid (C24),cerotic acid (C26), montanic acid (C28), and melissic acid (C30),including branched and substituted derivatives thereof.

Additional examples of unsaturated fatty acids that can be present inthe disclosed compositions include, but are not limited to, allenic andacetylenic acids, such as, C14: 2, 4, 5; C18: 5, 6 (laballenic); 5, 6,16 (lamenallenic); CIS: 6a (tarinic); 9a; 9a, 11t (ximenynic); 9a, 11a;9a, 11a, 13c (bolekic); 9a, 11a, 13a, 15; 8a, 10t (pyrulic) 9; 12a(crepenynic); 9c, 12a, 14c (dehydrocrepenynic acid); 6a, 9c, 12c; 6a,9c, 12c, 15c, 8a, 11c, 14c and corresponding Δ17e derivatives, 8-OHderivatives, and Δ17e, 8-OH derivatives. Branched-chain acids,particularly iso-acids and anteiso acids, polymethyl branched acids,phytol based acids (e.g., phytanic, pristanic), furanoid acids are alsosuitable fatty acids, including the residues derived therefrom, that canbe present in the disclosed compositions. Still further fatty acids andresidues thereof include, but are not limited to, cyclic acids, such ascyclopropane fatty acids, cyclopropene acids (e.g., lactobacillic),sterulic, malvalic, sterculynic, 2-hydroxysterculic, aleprolic,alepramic, aleprestic, aleprylic alepric, hydnocarpic, chaulmoogrichormelic, manaoic, gorlic, oncobic, cyclopentenyl acids, andcyclohexylalkanoic acids. Hydroxy acids, particularly butolic,ricinoleic, isoricinoleic, densipolic, lesquerolic, and auriolic arealso suitable fatty acids that can be present in the disclosedcompositions. Epoxy acids, particularly epoxidated C18:1 and C18:2, andfuranoid acids are further examples of fatty acids that can be presentin the disclosed compositions.

Trans Fatty Acids

In still other examples, the disclosed compositions can also containvarious amounts of trans-fatty acids. Trans-fatty acids are unsaturatedfatty acids where at least one double bond is a trans-double bond. Theamount of trans-fatty acids in the disclosed compositions can be lessthan or equal to about 5 wt. % based on the total weight of thecomposition. In other examples, the amount of trans-fatty acids can beless than or equal to about 5.0, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2,4.1, 4.0, 3.9, 3.8, 3.7, 3.6, 3.5, 3.4, 3.3, 3.2, 3.1, 3.0, 2.9, 2.8,2.7, 2.6, 2.5, 2.4, 2.3, 2.2, 2.1, 2.0, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4,1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1 wt. %based on the total weight of the composition, where any of the statedvalues can form an upper and/or lower endpoint as appropriate. Theamount of trans fatty acids can also be about 0 wt. %.

In further examples, the disclosed compositions can contain from about5.0 to about 0.0, from about 4.5 to about 0.0, from about 4.0 to about0.0, from about 3.5 to about 0.0, from about 3.0 to about 0.0, fromabout 2.5 to about 0.0, from about 2.0 to about 0.0, from about 1.5 toabout 0.0, from about 1.0 to about 0.0, from about 0.5 to about 0.0,from about 5.0 to about 0.5, from about 4.5 to about 0.5, from about 4.0to about 0.5, from about 3.5 to about 0.5, from about 3.0 to about 0.5,from about 2.5 to about 0.5, from about 2.0 to about 0.5, from about 1.5to about 0.5, from about 1.0 to about 0.5, from about 5.0 to about 1.0,from about 4.5 to about 1.0, from about 4.0 to about 1.0, from about 3.5to about 1.0, from about 3.0 to about 1.0, from about 2.5 to about 1.0,from about 2.0 to about 1.0, from about 1.5 to about 1.0, from about 5.0to about 2.0, from about 4.5 to about 2.0, from about 4.0 to about 2.0,from about 3.5 to about 2.0, from about 3.0 to about 2.0, from about 2.5to about 2.0, from about 5.0 to about 2.5, from about 4.5 to about 2.5,from about 4.0 to about 2.5, from about 3.5 to about 2.5, from about 3.0to about 2.5, from about 5.0 to about 3.0, from about 4.5 to about 3.0,from about 4.0 to about 3.0, from about 3.5 to about 3.0, from about 5.0to about 3.5, from about 4.5 to about 3.5, from about 4.0 to about 3.5,from about 5.0 to about 4.0, from about 4.5 to about 4.0, from about 5.0to about 4.5, wt. % of trans-fatty acids, based on the total weight ofthe composition.

Oxidized Fatty Acids

The disclosed compositions can also contain low levels of oxidized fattyacids and/or aldehydes. For example, many oils containing unsaturatedfatty acids can oxidize and break down, resulting in the production ofvolatile aldehydes like hexanal and a non-volatile portion of theoxidized fatty acid, which remains a part of the composition. Thecompositions disclosed herein can have reduced levels of such oxidizedfatty acids and/or aldehydes, as compared to other marina oils.

The amount of the aldehydes and oxidized fatty acids in an oil can bemeasured by reacting the oil with p-Anisidine and determining thep-Anisidine value. The p-Anisidine value is defined as 100 times theabsorbance (at 350 nm) of a solution resulting from reaction of 1 gramof oil in 100 mL of solvent. p-Anisidine values can be determined bywell known methods such as the method described in ISO method number6885:1998, which is available from the International Organization forStandardization; this method is incorporated by reference herein in itsentirety.

Typical p-Anisidine values for crude marine oil can be about 20. Thecompositions disclosed herein, however, can have p-Anisidine values ofless than or equal to about 25. In other examples, the disclosedcompositions can have p-Anisidine values of less than or equal to about25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7,6, 5, 4, 3, 2, or 1 as determined by ISO 6885:1998, where any of thestated values can form an upper and/or lower endpoint as appropriate.

In other examples of the disclosed compositions, the p-Anisidine valuecan be from about 25 to about 1, from about 24 to about 1, from about 23to about 1, from about 22 to about 1, from about 21 to about 1, fromabout 20 to about 1, from about 19 to about from about 18 to about 1,from about 17 to about 1, from about 16 to about 1, from about 15 toabout 1, from about 14 to about 1, from but 13 to about 1, from about 12to about 1, froze about 11 to about 1, from about 10 to about 1, fromabout 9 to about 1, from about 8 to about 1, from about 7 to about 1,from about 6 to about 1, from about 5 to about 1, from about 4 to about1, from about 3 to about 1, from about 2 to about 1, from about 25 toabout 2, from about 24 to about 2, from about 23 to about 2, from about22 to about 2, from about 21 to about 2, from about 20 to about 2, fromabout 19 to about 2, from about 18 to about 2, from about 17 to about 2,from about 16 to about 2, from about 15 to about 2, from about 14 toabout 2, from about 13 to about 2, from about 12 to about 2, from about11 to about 2, from about 10 to about 2, from about 9 to about 2, fromabout 8 to about 2, from about 7 to about 2, from about 6 to about 2,from about 5 to about 2, from about 4 to about 2, from about 3 to about2, from about 25 to about 3, from about 24 to about 3, from about 23 toabout 3, from about 22 to about 3, from about 21 to about 3, from about20 to about 3, from about 19 to about 3, from about 18 to about 3, fromabout 17 to about 3, from about 16 to about 3, from about 1.5 to about3, from about 14 to about 3, from about 13 to about 3, from about 12 toabout 3, from about 11 to about 3, from about 10 to about 3, from about9 to about 3, from about 8 to about 3, from about 7 to about 3, fromabout 6 to about 3, from about 5 to about 3, from about 4 to about 3,from about 25 to about 4, from about 24 to about 4, from about 23 toabout 4, from about 22 to about 4, from about 21 to about 4, from about20 to about 4, from about 19 to about 4, from about 18 to about 4, fromabout 17 to about 4, from about 16 to about 4, from about 15 to about 4,from about 14 to about 4, from about 13 to about 4, from about 12 toabout 4, from about 11 to about 4, from about 10 to about 4, from about9 to about 4, from about 8 to about 4, from about 7 to about 4, fromabout 6 to about 4, from about 5 to about 4, from about 25 to about 5,from about 24 to about 5, from about 23 to about 5, from about 22 toabout 5, from about 21 to about 5, from about 20 to about 5, from about19 to about 5, from about 18 to about 5, from about 17 to about 5, fromabout 16 to about 5, from about 15 to about 5, from about 14 to about 5,from about 13 to about 5, from about 12 to about 5, from about 11 toabout 5, from about 10 to about 5, from about 9 to about 5, from about 8to about 5, from about 7 to about 5, from about 6 to about 5, from about25 to about 6, from about 24 to about 6, from about 23 to about 6, fromabout 22 to about 6, from about 21 to about 6, from about 20 to about 6,from about 19 to about 6, from about 18 to about 6, from about 17 toabout 6, from about 16 to about 6, from about 15 to about 6, from about14 to about 6, from about 13 to about 6, from about 12 to about 6, fromabout 11, to about 6, from about 10 to about 6, from about 9 to about 6,from about 8 to about 6, from about 7 to about 6, from about 25 to about7, from about 24 to about 7, from about 23 to about 7, from about 22 toabout 7, from about 21 to about 7, from about 20 to about 7, from about19 to about 7, from about 18 to about 7, from about 17 to about 7, fromabout 16 to about 7, from about 15 to about 7, from about 14 to about 7,from about 13 to about 7, from about 12 to about 7, from about 11 toabout 7, from about 10 to about 7, from about 9 to about 7, from about 8to about 7, from about 25 to about 8, from about 24 to about 8, fromabout 23 to about 8, from about 22 to about 8, from about 21 to about 8,from about 20 to about 8, from about 19 to about 8, from about 18 toabout 8, from about 17 to about 8, from about 16 to about 8, from about15 to about 8, from about 14 to about 8, from about 13 to about 8, fromabout 12 to about 8, from about 11 to about 8, from about 10 to about 8,from about 9 to about 8, from about 25 to about 9, from about 24 toabout 9, from about 23 to about 9, from about 22 to about 9, from about21 to about 9, from about 20 to about 9, from about 19 to about 9, fromabout 18 to about 9, from about 17 to about 9, from about 16 to about 9,from about 15 to about 9, from about 14 to about 9, from about 13 toabout 9, from about 12 to about 9, from about 11 to about 9, from about10 to about 9, from about 25 to about 10, from about 24 to about 10,from about 23 to about 10, from about 22 to about 10, from about 21 toabout 10, from about 20 to about 10, from about 19 to about 10, fromabout 18 to about 10, from about 17 to about 10, from about 16 to about10, from about 15 to about 10, from about 14 to about 10, from about 13to about 10, from about 12 to about 10, from about 11 to about 10, fromabout 25 to about 11, from about 24 to about 11, from about 23 to about11, from about 22 to about 11, from about 21 to about 11, from about 20to about 11, from about 19 to about 11, from about 18 to about 11, fromabout 17 to about 11, from about 16 to about 11, from about 15 to about11, from about 14 to about 11, from about 13 to about 11, from about 12to about 11, from about 25 to about 12, from about 24 to about 12, fromabout 23 to about 12, from about 22 to about 12, from about 21 to about12, from about 20 to about 12, from about 19 to about 12, from about 18to about 12, from about 17 to about 12, from about 16 to about 12, fromabout 15 to about 12, from about 14 to about 12, from about 13 to about12, from about 25 to about 13, from about 24 to about 13, from about 23to about 13, from about 22 to about 13, from about 21 to about 13, fromabout 20 to about 13, from about 19 to about 13, from about 18 to about13, from about 17 to about 13, from about 16 to about 13, from about 15to about 13, from about 14 to about 13, from about 25 to about 14, fromabout 24 to about 14, from about 23 to about 14, from about 22 to about14, from about 21 to about 14, from about 20 to about 14, from about 19to about 14, from about 18 to about 14, from about 17 to about 14, fromabout 16 to about 14, from about 15 to about 14, from about 25 to about15, from about 24 to about 15, from about 23 to about 15, from about 22to about 15, from about 21 to about 15, from about 20 to about 15, fromabout 19 to about 15, from about 18 to about 15, from about 17 to about15, from about 16 to about 15, from about 25 to about 16, from about 24to about 16, from about 23 to about 16, from about 22 to about 16, fromabout 21 to about 16, from about 20 to about 16, from about 19 to about16, from about 18 to about 16, from about 17 to about 16, from about 23to about 17, from about 24 to about 17, from about 23 to about 17, fromabout 22 to about 17, from about 21 to about 17, from about 20 to about17, from about 19 to about 17, from about 18 to about 17, from about 25to about 18, from about 24 to about 18, from about 23 to about 18, fromabout 22 to about 18, from about 21 to about 18, from about 20 to about18, from about 19 to about 18, from about 25 to about 19, from about 24to about 19, from about 23 to about 19, from about 22 to about 19, fromabout 21 to about 19, from about 20 to about 19, from about 25 to about20, from about 24 to about 20, from about 23 to about 20, from about 22to about 20, from about 21 to about 20, from about 25 to about 21, fromabout 24 to about 21, from about 23 to about 21, from about 22 to about21, from about 25 to about 22, from about 24 to about 22, from about 23to about 22, from about 25 to about 23, from about 24 to about 23, orfrom about 25 to about 24 as determined by ISO 6885:1998.

Levels of Mono-, Di-, and Tri-Glycerides

The disclosed compositions can also comprise high levels oftriglycerides. Triglycerides are esters of fatty acids, such as thosedisclosed herein, and a tri-functional alcohol, i.e., 1,2,3-propantriol,which is also known as glycerol. In many examples, the disclosedcompositions can comprise greater than or equal to about 97 wt. %triglycerides based on the total weight of the composition. In otherexamples, the disclosed compositions can comprise about 97.5, 98.0,98.5, 99.0, 99.3, 99.7, 99.9, or 100 wt. % triglycerides based on thetotal weight of the composition, where any of the stated values can forman upper and/or lower endpoint when appropriate. In still otherexamples, the disclosed compositions can comprise from about 97.0 toabout 100.0, from about 97.5 to about 100.0, from about 98.0 to about100.0, from about 98.5 to about 100.0, from about 99.0 to about 100.0,from about 99.5 to about 100.0, from about 99.7 to about 100.0, fromabout 99.9 to about 100.0, from about 97.0 to about 99.9, from about97.5 to about 99.9, from about 98.0 to about 99.9, from about 98.5 toabout 99.9, from about 99.0 to about 99.9, from about 99.5 to about99.9, from about 99.7 to about 99.9, from about 97.0 to about 99.7, fromabout 97.5 to about 99.2, from about 98.0 to about 99.7, from about 98.5to about 99.7, from about 99.0 to about 99.7, from about 99.5 to about99.7, from about 97.0 to about 99.5, from about 97.5 to about 99.5, fromabout 98.0 to about 99.5, from about 98.5 to about 99.5, from about 99.0to about 99.5, from about 97.0 to about 99.0, from about 97.5 to about99.0, from about 98.0 to about 99.0, from about 98.5 to about 99.0, fromabout 97.0 to about 98.5, from about 97.5 to about 98.5, from about 98.0to about 98.5, from about 97.0 to about 98.0, from about 97.5 to about98.0, from about 97.0 to about 97.5 wt. % triglycerides based on thetotal weight of the composition.

Also, the disclosed compositions can comprise loss than or equal toabout 2 wt. % di- and/or mono-glycerides based on the total weight ofthe composition. For example, the disclosed compositions can compriseless than or equal to about 1.5, 1.0, 0.5, or 0.0 wt. % di- and/ormono-glycerides based on the total weight of the composition, where anyof the stated values can form an upper and/or lower endpoint asappropriate. In a particular example, the disclosed compositions cancomprise 0.0 wt. % monoglycerides.

Methods of Making

The disclosed compositions can be prepared by methods disclosed herein.That is, the disclosed methods can be used to reduce and/or removecompounds (e.g., sterols) from oils (e.g., marine oils, plant oils, andanimal oils), producing the disclosed compositions. In one aspect, thedisclosed methods comprise contacting an oil with an adsorbent toprovide a mixture, heating the mixture to from about 100 to about 210°C., and removing the adsorbent from the mixture, to provide thedisclosed composition. In a specific example, disclosed herein is amethod for preparing a low-cholesterol composition by contacting an oilwith an adsorbent to provide a mixture, heating the mixture to fromabout 100 to about 210° C., and removing the adsorbent from the mixture.These disclosed methods can be used to provide a low-cholesterolcomposition wherein the composition comprises, for example, less thanabout 2.0 milligrams of cholesterol per gram of the composition.

As noted, the resulting composition can comprise reduced levels ofcompounds such as sterols, as is described above. Further, the resultingcompositions can comprise levels of EPA/DHA, Gardner color, p-Anisidinevalues, trans-fatty acid levels, and/or mono-, di-, and tri-glyceridelevels as described above.

Adsorbent

In the disclosed methods, the adsorbent can be silica, clay, carbon, ora mixture thereof. Suitable examples of silica can be TRYSIL™, which isavailable from Grace Davison (Columbia Md.). Suitable examples of claycan be Grade F-160 activated clay, which is available from manufacturedby Englehardt Corporation (Jackson, Miss.).

In some examples of the disclosed methods, the adsorbent can be acombination of silica and clay. Suitable combinations can comprisesilica and clay in a weight ratio of about 1:1. In other examples, theadsorbent can comprise silica and clay in a weight ratio of about 1:10,1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1,7:1, 8:1, 9:1, or 10:1.

In the disclosed methods, the amount of adsorbent used can be from lessthan or equal to about 20 wt. %, based on the weight of the oil. Forexample, the adsorbent can used in an amount of about 20, 19, 18, 17,16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 wt. %, based onthe weight of the oil, where any of the stated values can form an upperand/or lower endpoint as appropriate. In other examples, the adsorbentcan be used in an amount of from about 20 to about 2, from about 18 toabout 2, from about 16 to about 2, from about 14 to about 2, from about12 to about 2, from about 10 to about 2, from about 8 to about 2, fromabout 6 to about 2, from about 4 to about 2, from about 20 to about 4,from about 18 to about 4, from about 16 to about 4, from about 14 toabout 4, from about 12 to about 4, from about 10 to about 4, from about8 to about 4, from about 6 to about 4, from about 20 to about 6, fromabout 18 to about 6, from about 16 to about 6, from about 14 to about 6,from about 12 to about 6, from about 10 to about 6, from about 8 toabout 6, from about 20 to about 8, from about 18 to about 8, from about16 to about 8, from about 14 to about 8, from about 12 to about 8, fromabout 10 to about 8, from about 20 to about 10, from about 18 to about10, from about 16 to about 10, from about 14 to about 10, from about 12to about 10, from about 20 to about 12, from about 18 to about 12, fromabout 16 to about 12, from about 14 to about 12, from about 20 to about14, from about 18 to about 14, from about 16 to about 14, from about 20to about 16, from about 18 to about 16, or from about 20 to about 18 wt.%, based on the weight of the oil.

Also, the ratio of adsorbent to oil that can be used can be about20:100, 19:100, 18:100, 17:100, 16:100, 15:100, 14:100, 13:100, 12:100,11:100, 1:10, 9:100, 8:100, 7:100, 6:100, 5:100, 4:100, 3:100, 2:100, or1:100. When the adsorbent is a combination of silica and clay, theindividual ratios of silica and clay to oil that can be used can beabout 1:10, 9:100, 8:100, 7:100, 6:100, 5:100, 4:100, 3:100, 2:100, or1:100. Also, when the adsorbent is a combination of silica and clay,each can be used in an amount of about 20, 19, 18, 17, 16, 15, 14, 13,12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 wt. %, based on the weight ofthe oil, where any of the stated values can form an upper and/or lowerendpoint as appropriate. In several examples, from about 3 to about 7wt. % (e.g., or from about 5 to about 7 wt. %) of the adsorbent iscontacted to the oil.

Mixing

The adsorbent and oil can be mixed by any methods blown in the art.“Mixing” is not meant to imply a particular outcome of mixing, such asthe dissolution of any components to a particular level or the formationof a particular composition, such as homogeneous mixture, although suchmixtures can be produced and some components can be dissolved by mixing.It can be desired that the mixing be vigorous. Mixing can be performedmanually or by a mechanical mixing device such as, but not limited to, astatic mixer, a magnetic stirrer, a shaker, spinner, or rotating device.Mixing can also be performed by forcing or bubbling a gas through themixture or by sonication.

Mixing the oil and adsorbent can be performed for at least 1 minute.Mixing can also be performed for at least 1, 5, 15, 20, 25, 30, 35, 40,45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 minutes, where any ofthe stated values can form an upper and/or lower endpoint asappropriate. Also, the mixture can be mixed before, during, or after theheating step.

Temperature

Heating the mixture of adsorbent and oil can be performed at varioustemperatures, but, typically, the method can take place at an elevatedtemperature. The precise elevated temperature can depend on theparticular oil and amount thereof being used, the particular adsorbentand the amount thereof being used, the ratio of oil to the adsorbent,the particular pressure, preference, and the like. Suitable temperaturesat which the disclosed methods can be performed include, but are notlimited to, from about 100 to about 210° C., from about 110 to about200° C., from about 120 to about 190° C., from about 130 to about 180°C., from about 140 to about 170° C., or from about 150 to about 160° C.In other examples, the mixture can be heated to about 100, 101, 102,103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116,117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130,131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144,145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158,159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172,173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186,187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200,201, 202, 203, 204, 205, 206, 207, 208, 209, or 210° C., where any ofthe stated values can form an upper and/or lower endpoint asappropriate.

In yet other examples, the mixture can be heated to from about 100 toabout 210, from about 110 to about 210, from about 120 to about 210,from about 130 to about 210, from about 140 to about 210, from about 150to about 210, from about 160 to about 210, from about 170 to about 210,from about 180 to about 210, from about 190 to about 210, from about 200to about 210, from about 100 to about 200, from about 110 to about 200,from about 120 to about 200, from about 130 to about 200, from about 140to about 200, from about 150 to about 200, from about 160 to about 200,from about 170 to about 200, from about 180 to about 200, from about 190to about 200, from about 100 to about 190, from about 110 to about 190,from about 130 to about 190, from about 140 to about 190, from about 150to about 190, from about 160 to about 190, from about 170 to about 190,from about 180 to about 190, from about 100 to about 180, from about 110to about 180, from about 120 to about 180, from about 140 to about 180,from about 150 to about 180, from about 160 to about 180, from about 170to about 180, from about 100 to about 170, from about 110 to about 170,from about 120 to about 170, from about 130 to about 170, from about 150to about 170, from about 160 to about 170, from about 100 to about 160,from about 110 to about 160, from about 120 to about 160, from about 130to about 160, from about 140 to about 160, from about 100 to about 150,from about 110 to about 150, from about 120 to about 150, from about 130to about 150, from about 140 to about 150, from about 100 to about 140,from about 110 to about 140, from about 120 to about 140, from about 130to about 140, from about 100 to about 130, from about 110 to about 130,from about 120 to about 130, from about 100 to about 120, from about 110to about 120, or from about 100 to about 110° C. In some specificexamples, the mixture can be heated to from about 140 to about 180° C.or, more specifically, from about 180 to about 190° C. or from about 150to about 170° C.

It is also contemplated that the oil is heated prior to contacting withthe adsorbent. Such a pre-heating step can be performed at any oftemperatures and temperature ranges described herein.

Heating the mixture and/or pre-heating the oil can take place over aperiod of time, for example for at least 1, 10, 20, 30, 40, 50, or 60minutes. In some examples, the heating step is performed far from about10 to about 20, from about 20 to about 30, from about 10 to about 30, orfrom about 30 to about 60 minutes. Further, after heating, the mixturecan be allowed to cool from about 30 to about 60 minutes.

Pressure

In the disclosed methods, the heating step can be conducted underreduced pressure. A suitable pressure is less than or equal to about 1Torr or less than or equal to about 0.1 Torr. In other examples, theheating step can be conducted at a pressure of less than or equal toabout 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07,0.06, 0.05, 0.04, 0.03, 0.02, or 0.01, where any of the stated valuescan form an upper and/or lower endpoint when appropriate.

Separation

After the mixture is heated, the adsorbent can be removed from themixture. Removing the adsorbent can be performed by any methods known inthe art. For example, the adsorbent can be removed by filtration,centrifugation, or other like methods. The compositions that result fromheating the oil with the adsorbent, and also from removing theadsorbent, can comprise a reduced content of cholesterol (e.g., lessthan or equal to about 6 mg of cholesterol per gram of oil), ap-Anisidine value of less than or equal to about 25, a Gardner color ofless than or equal to about 18, from about 0 to about 700 milligrams ofDHA and/or EPA per gram of the composition (or a EPA:DHA wt. % ratio offrom 0:70 to 70:0), less than or equal to about 5 wt. % trans-fattyacids, and/or greater than or equal to about 97 wt. % triglycerides. Thecomposition obtained includes cholesterol-free product. The process alsocan be used as bleaching process for a dual purpose, i.e. to reducecholesterol and color from oil.

Supplements

Also, disclosed herein are nutritional supplements comprising thecompositions disclosed herein. A nutritional supplement is any compoundor composition that can be administered to or taken by a subject toprovide, supply, or increase a nutrient(s) (e.g., vitamin, mineral,essential trace element, amino acid, peptide, nucleic acid,oligonucleotide, lipid, cholesterol, steroid, carbohydrate, and thelike). In one aspect, disclosed herein are nutritional supplementscomprising any of the compounds disclosed herein. For example, anutritional supplement can comprise a composition comprising a marineoil, wherein the composition has less than or equal to about 6 mg ofcholesterol per gram of oil, a p-Anisidine value of less than or equalto 25, a Gardner color of less than or equal to 18, from about 0 toabout 700 milligrams of DHA and/or EPA per gram of the composition (or aEPA:DHA wt. % ratio of from 0:70 to 70:0), less or equal to about 5 wt.% trans-fatty acids, and/or greater than or equal to about 97 wt. %triglycerides.

The nutritional supplement can comprise any amount of the compositionsdisclosed herein, but will typically contain an amount determined tosupply a subject with a desired dose of an oil or particular fatty acid(e.g., EPA and/or DHA). The exact amount of compound required in thenutritional supplement will vary from subject to subject, depending onthe species, age, weight and general condition of the subject, theseverity of any dietary deficiency being treated, the particular mode ofadministration, and the like. Thus, it is not possible to specify anexact amount for every nutritional supplement. However, an appropriateamount can be determined by one of ordinary skill in the art using onlyroutine experimentation given the teachings herein.

The nutritional supplement can also comprise other nutrient(s) such asvitamins other trace elements, minerals, and the like. Further, thenutritional supplement can comprise other components such aspreservatives, antimicrobials, anti-oxidants, chelating agents,thickeners, flavorings, diluents, emulsifiers, dispersing aids, orbinders.

The nutritional supplements are generally taken orally and can be in anyform suitable for oral administration. For example, a nutritionalsupplement can typically be in a tablet, gel-cap, capsule, liquid,sachets, or syrup form.

Pharmaceutical Formulations

Also, disclosed herein are pharmaceutical formulations comprising thecompositions disclosed herein. A suitable pharmaceutical formulation cancomprise any of the disclosed compositions with a pharmaceuticallyacceptable carrier. For example, a pharmaceutical formulation cancomprise composition comprising an oil (e.g., marine oil), wherein thecomposition has less than or equal to about 6 mg of cholesterol per gramof oil, a p-Anisidine value of less than or equal to 25, a Gardner colorof less than or equal to 18, from about 0 to about 700 milligrams of DHAand/or EPA per gram of the composition (or a EPA:DHA wt. % ratio of from0:70 to 70:0), lass or equal to about 5 wt. % trans-fatty acids, and/orgreater than or equal to about 97 wt. % triglycerides, and apharmaceutically acceptable carrier. The disclosed pharmaceuticalformulations can be used therapeutically or prophylactically.

By “pharmaceutically acceptable” is meant a material that is notbiologically or otherwise undesirable, i.e., the material may beadministered to a subject without causing any undesirable biologicaleffects or interacting in a deleterious manner with any of the othercomponents of the pharmaceutical formulation in which it is contained.The carrier would naturally be selected to minimize any degradation ofthe active ingredient and to minimize any adverse side effects in thesubject, as would be well known to one of skill in the art.

Pharmaceutical carriers are known to those stilled in the art. Thesemost typically would be standard carriers for administration of drugs tohumans, including solutions such as sterile water, saline, and bufferedsolutions at physiological pH. Suitable carriers and their formulationsare described in Remington: The Science and Practice of Pharmacy,21^(st) ed., Lippincott Williams & Wilkins, Philadelphia, Pa., 2005,which is incorporated by reference herein for its teachings of carriersand pharmaceutical formulations. Typically, an appropriate amount of apharmaceutically-acceptable salt is used in the formulation to renderthe formulation isotonic. Examples of the pharmaceutically-acceptablecarrier include, but are not limited to, saline, Ringer's solution anddextrose solution. The pH of the solution can be from about 5 to about 8(e.g., from about 7 to about 7.5). Further carriers include sustainedrelease preparations such as semipermeable matrices of solid hydrophobicpolymers containing the disclosed compounds, which matrices are in theform of shaped articles, e.g., films, liposomes, microparticles, ormicrocapsules. It will be apparent to those persons skilled in the artthat certain carriers can be more preferable depending upon, forinstance, the route of administration and concentration of compositionbeing administered. Other compounds can be administered according tostandard procedures used by those skilled in the art.

Pharmaceutical formulations can include additional carriers, as well asthickeners, diluents, buffers, preservatives, surface active agents andthe like in addition to the compounds disclosed herein. Pharmaceuticalformulations can also include one or more additional active ingredientssuch as antimicrobial agents, anti-inflammatory agents, anesthetics, andthe like.

The pharmaceutical formulation can be administered in a number of waysdepending on whether local or systemic treatment is desired, and on thearea to be treated. Administration can be topically (includingophthalmically, vaginally, rectally, intranasally), orally, byinhalation, or parenterally, for example by intravenous drip,subcutaneous, intraperitoneal or intramuscular injection. The disclosedcompounds can be administered intravenously, intraperitoneally,intramuscularly, subcutaneously, intracavity, or transdermally.

Preparations for parenteral administration include sterile aqueous ornon-aqueous solutions, suspensions, and emulsions. Examples ofnon-aqueous solvents are propylene glycol, polyethylene glycol,vegetable oils such as olive oil, marine oils, and injectable organicesters such as ethyl oleate. Aqueous carriers include water,alcoholic/aqueous solutions, and emulsions or suspensions, includingsaline and buffered media. Parenteral vehicles include sodium chloridesolution, Ringer's dextrose, dextrose and sodium chloride, lactatedRinger's, and fixed oils. Intravenous vehicles include fluid andnutrient replenishers, electrolyte replenishers (such as those based onRinger's dextrose), and the like. Preservatives and other additives mayalso be present such as, for example, antimicrobials, anti-oxidants,chelating agents, and inert gases and the like.

Pharmaceutical formulations for topical administration may includeointments, lotions, creams, gels, drops, suppositories, sprays, liquidsand powders. Conventional pharmaceutical carriers, aqueous, powder oroily bases, thickeners and the like can be desirable.

Pharmaceutical formulations for oral administration include, but are notlimited to, powders or granules, suspensions or solutions in water ornon-aqueous media, capsules, sachets, or tablets. Thickeners,flavorings, diluents, emulsifiers, dispersing aids, or binders can bedesirable.

Some of the formulations can potentially be administered as apharmaceutically acceptable acid- or base-addition salt, formed byreaction with inorganic acids such as hydrochloric acid, hydrobromicacid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, andphosphoric acid, and organic acids such as formic acid, acetic acid,propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid,malonic acid, succinic acid, maleic acid, and fumaric acid, or byreaction with an inorganic base such as sodium hydroxide, ammoniumhydroxide, potassium hydroxide, and organic bases such as mono-, di-,trialkyl and aryl amines and substituted ethanolamines.

Delivery Devices

Any of the compositions described herein can be incorporated into adelivery device. Examples of delivery devices include, but are notlimited to, microcapsules, microspheres, nanospheres or nanoparticles,liposomes, noisome, nanoerythrosome, solid-liquid nanoparticles, gels,gel capsules, tablets, lotions, creams, sprays, emulsions, or powders.Other examples of delivery devices that are suitable for non-oraladministration include pulmospheres. Examples of particular deliverydevices useful herein are described below.

The disclosed compounds can be incorporated into liposomes. As is knownin the art, liposome are generally derived from phospholipids or otherlipid substances. Liposomes are formed by mono- or multi-lamellarhydrated liquid crystals that are dispersed in an aqueous medium. Anynon-toxic, physiologically acceptable and metabolizable lipid capable offorming liposomes can be used. The disclosed compositions in liposomeform can contain, in addition to a compositions disclosed herein,stabilizers, preservatives, excipients, and the like. Examples ofsuitable lipids are the phospholipids and the phosphatidyl cholines(lecithins), both natural and synthetic. Methods of forming liposomesare known in the art. See, e.g., Prescott, Ed., Methods in Cell Biology,Volume XIV, Academic Press, New York, p. 33 et seq., 1976, which ishereby incorporated by reference herein for its teachings of liposomesand their preparation.

In other examples, the liposomes can be cationic liposomes DOTMA, POPE,DC cholesterol) or anionic liposomes. Liposomes can further compriseproteins to facilitate targeting a particular cell, if desired.Administration of a composition comprising a compound and a cationicliposome can be administered to the blood afferent to a target organ orinhaled into the respiratory tract to target cells of the respiratorytract. Regarding liposomes, see e.g., Brigham et al., Am. J. Resp. Cell.Mol. Biol. 1989, 1:95-100; Felgner et al., Proc. Natl. Acad. Sci. USA1987, 84:7413-7; and U.S. Pat. No. 4,897,355, which are incorporated byreference herein for their teachings of liposomes. As one example,delivery can be via a liposome using commercially available liposomepreparations such as LIPOFECTIN, LIPOFECTAMINE (GIBCO-BRL, Inc.,Gaithersburg, Md.), SUPERFECT (Qiagen, Inc. Hilden, Germany) andTRANSFECTAM (Promega Biotec, Inc., Madison, Wis.), as well as otherliposomes developed according to procedures standard in the art.Liposomes where the diffusion of the compound or delivery of thecompound from the liposome is designed for a specific rate or dosage canalso be used.

As described herein, niosomes are delivery devices that can be used todeliver the disclosed compositions. Noisomes are multilamellar orunilamellar vesicles involving non-ionic surfactants. An aqueoussolution of solute is enclosed by a bilayer resulting from theorganization of surfactant macromolecules. Similar to liposomes,noisomes are used in targeted delivery of for example, anticancer drugs,including methotrexate, doxorubicin, and immunoadjuvants. They aregenerally understood to be different from transferosomes, vesiclesprepared from amphiphilic carbohydrate and amino group containingpolymers, e.g., chitosan.

As described herein, nanoerythrosomes are delivery devices that can beused to deliver the disclosed compositions. Nanoerythrosomes arenano-vesicles made of red blood cells via dialysis through filters ofdefined pore size. These vesicles can be loaded with a diverse-array ofbiologically active molecules, including proteins and the compositionsdisclosed herein. They generally serve as ideal carriers forantineoplastic agents like bleomycin, actinomycin D, but can be used forsteroids, other lipids, etc.

Artificial red blood cells are further delivery devices that can be usedto deliver the disclosed compositions. Artificial red blood cells can begenerated by interfacial polymerization and complex emulsion methods.Generally, the “cell” wall is made of polyphtaloyl L-lysinepolymer/polystyrene and the core is made of a hemoglobin solution fromsheep hemolysate. Hemoglobin loaded microspheres typically have particlesizes of from about 1 to about 10 mm. Their size, flexibility, andoxygen carrying capacity is similar to red blood cells.

Solid-lipid nanoparticles are other delivery devices that can be used todeliver the disclosed compositions. Solid-lipid nanoparticles arenanoparticles that are dispersed in an aqueous surfactant solution. Theyare comprised of a solid hydrophobic core having a monolayer of aphospholipid coating and are usually prepared by high-pressurehomogenization techniques. Immunomodulating complexes (ISCOMS) areexamples of solid-lipid nanoparticles. They are cage-like 40 nmsupramolecular assemblies comprising of phospholipid, cholesterol, andhydrophobic antigens and are used mostly as immunoadjuvants. Forinstance, ISCOMs are used to prolong blood-plasma levels ofsubcutaneously injected cyclosporine.

Microspheres and micro-capsules are yet other delivery devices that canbe used to deliver the disclosed compositions. In contrast to liposomaldelivery systems, microspheres and micro-capsules typically do not havean aqueous core but a solid polymer matrix or membrane. These deliverydevices are obtained by controlled precipitation of polymers, chemicalcross-linking of soluble polymers, and interfacial polymerization of twomonomers or high-pressure homogenization techniques. The encapsulatedcompound is gradually released from the depot by erosion or diffusionfrom the particles. Successful formulations of short acting peptides,such as LHRH agonists like leuprorelin and triptoreline, have beendeveloped. Poly(lactide co-glycolide (PLGA) microspheres are currentlyused as monthly and three monthly dosage forms in the treatment ofadvanced prostrate cancer, endometriosis, and other hormone responsiveconditions. Leuprolide, an LHRH superagonist, was incorporated into avariety of PLGA matrices using a solvent extraction/evaporation method.As noted, all of these delivery devices can be used with the disclosedcompositions.

Pulmospheres are still other examples of delivery devices that can beused herein. Pulmospheres are hollow porous particles with a low density(less than about 0.1 g/mL). Pulmospheres typically have excellentre-dispersibility and are usually prepared by supercritical fluidcondensation technology. Co-spray-drying with certain matrices, such ascarbohydrates, human serum albumin, etc., can improve the stability ofproteins and peptides (e.g., insulin) and other biomolecules forpulmonary delivery. This type of delivery could be also accomplishedwith micro-emulsions and lipid emulsions, which are ultra fine, thin,transparent oil-in-water (o/w) emulsions formed spontaneously with nosignificant input of mechanical energy. In this technique, an emulsioncan be prepared at a temperature, which must be higher than the phaseinversion temperature of the system. At elevated temperature theemulsion is of water-in-oil (w/o) type and as it cools at the phaseinversion temperature, this emulsion is inverted to become o/w. Due totheir very small inner phase, they are extremely stable and used forsustained release of steroids and vaccines. Lipid emulsions comprise aneutral lipid core (i.e., triglycerides) stabilized by a monolayer ofamphiphilic lipid (i.e., phospholipid) using surfactants like egglecithin triglycerides and miglyol. They are suitable for passive andactive targeting.

There are other oral delivery systems under investigation that are basedon osmotic pressure modulation, pH modulation, swelling modulation,altered density and floating systems, mucoadhesiveness etc. Theseformulations and time-delayed formulations to deliver drugs inaccordance with circadian rhythm of disease that are currently in use orinvestigation can be applied for delivery of the disclosed compositions.

Microcapsules

In one aspect disclosed herein, the disclosed compositions can beincorporated into microcapsules. For example, the microcapsule cancomprise an agglomeration of primary microcapsules and disclosedcompositions, each individual primary microcapsule having a primaryshell, wherein the disclosed compositions are encapsulated by theprimary shell, wherein the agglomeration is encapsulated by an outershell. These microcapsules are referred to herein as “multicoremicrocapsules.”

In another aspect, described herein are microcapsules comprising acomposition disclosed herein, a primary shell, and a secondary shell,wherein the primary shell encapsulates the disclosed composition, andthe secondary shell encapsulates the loading substance and primaryshell. These microcapsules are referred to herein as “single-coremicrocapsules.

Optionally, other loading substances can be encapsulated with thedisclosed compositions. This additional loading substance can be anysubstance that is not entirely soluble in the aqueous mixture. In oneaspect, the loading substance is a solid, a hydrophobic liquid, or amixture of a solid and a hydrophobic liquid. In another aspect, theloading substance comprises a grease, an oil, a lipid, a drug (e.g.,small molecule), a biologically active substance, a nutritionalsupplement (e.g., vitamins), a flavor compound, or a mixture thereof.Examples of oils include, but are not limited to, animal oils (e.g.,fish oil, marine mammal oil, etc.), vegetable oils (e.g., canola orrapeseed), mineral oils, derivatives thereof or mixtures thereof. Theloading substance can be a purified or partially purified oily substancesuch as a fatty acid, a triglyceride or ester thereof, or a mixturethereof. In another aspect, the loading substance can be a carotenoid(e.g., lycopene), a satiety agent, a flavor compound, a drug (e.g., awater insoluble drug), a particulate, an agricultural chemical (e.g.,herbicides, insecticides, fertilizers), or an aquaculture ingredient(e.g., feed, pigment).

In one aspect, the loading substance can be an omega-3 fatty acid, asdisclosed above, including derivatives thereof. Many types ofderivatives of omega-3 fatty acids are well known in the art. Examplesof suitable derivatives include, but are not limited to, esters, such asphytosterol esters, branched or unbranched C₁-C₃₀ alkyl esters, branchedor unbranched C₂-C₃₀ alkenyl esters, or branched or unbranched C₃-C₃₀cycloalkyl esters such as phytosterol esters and C₁-C₆ alkyl esters.Sources of oils can be derived from aquatic organisms (e.g., anchovies,capelin, Atlantic cod, Atlantic belying, Atlantic mackerel, Atlanticmenhaden, salmonids, sardines, shark, tuna, etc) and plants (e.g. flax,vegetables, etc) and microorganisms (e.g., fungi and algae).

In one aspect, the loading substance can contain an antioxidant.Examples of antioxidants include, but are not limited to, vitamin E,CoQ₁₀, tocopherols, lipid soluble derivatives of more polar antioxidantssuch as ascorbyl fatty acid esters (e.g., ascorbyl palmitate), plantextracts (e.g., rosemary, sage and oregano oils), algal extracts, andsynthetic antioxidants (e.g., BHT, TBHQ, ethoxyquin, alkyl gallates,hydroquinones, tocotrienols).

A number of different polymers can be used to produce the shell layersof the single and multicore microcapsules. Examples of such polymersinclude, but are not limited to, a protein, a polyphosphate, apolysaccharide, or a mixture thereof. In another aspect, the shellmaterial used to prepare the single- and multicore microcapsules furthercomprises In another aspect, the shell material used to prepare thesingle- and multicore microcapsules further comprises gelatin type A,gelatin type B, polyphosphate, gum arabic, alginate, chitosan,carrageenan, pectin, starch, modified starch, alfa-lactalbumin,beta-lactoglobumin, ovalbumin, polysorbiton, maltodextrins,cyclodextrins, cellulose, methyl cellulose, ethyl cellulose,hydropropylmethylcellulose, carboxymethylcellulose, milk protein, wheyprotein, soy protein, canola protein, albumin, chitin, polylactides,poly-lactide-co-glycolides, derivatized chitin, chitosan, poly-lysine,various inorganic-organic composites, or any mixture thereof. It is alsocontemplated that derivatives of these polymers can be used as well. Inanother aspect, the polymer can be kosher gelatin, non-kosher gelatin,Halal gelatin, or non-Halal gelatin.

In one aspect, one or more of the shell layers in the single andmulticore microcapsules comprises gelatin having a Bloom number lessthan 50. This gelatin is referred to herein as “low Bloom gelatin.” TheBloom number describes the gel strength formed at 10° C. with a 6.67%solution gelled for 18 hours. In one aspect, the low Bloom gelatin has aBloom number less than 40, less than 30, less than 20, or less than 10.In another aspect, the gelatin has a Bloom number of 45, 40, 35, 30, 25,20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0, where any stated value canfor an upper and/or lower endpoint as appropriate. In another aspect,the low Bloom gelatin is in both the primary shell and the outer shellof the multicore microcapsule. In one aspect, the low Bloom gelatin isgelatin type A. In another aspect, the low Bloom gelatin is gelatin typeA produced by Kenney & Ross Ltd., R.R. #3 Shelburne, NS Canada. Inanother aspect, gelatin having a Bloom number of zero is in both theprimary shell and the outer shell of the multicore microcapsule.

The material used to make the shells of the single- or multicoremicrocapsules can be a two-component system made from a mixture of twodifferent types of polymers. For example, the material can be a complexcoacervate between the polymer components. Complex coacervation iscaused by the interaction between two oppositely charged polymers. Inone aspect, the shell material used to produce the single and multicoremicrocapsules is composed of (1) low Bloom gelatin and (2) gelatin typeB, polyphosphate, gum arabic, alginate, chitosan, carrageenan, pectin,carboxymethylcellulose, whey protein, soy protein, canola protein,albumin, or a mixture thereof. The molar ratio of the different polymerscan vary. For example, the molar ratio of low Bloom gelatin to the otherpolymer component is from 1:5 to 15:1. For example, when low Bloomgelatin and polyphosphate are used, the molar ratio of low Bloom gelatinto polyphosphate is about 8:1 to about 12:1; when low Bloom gelatin andgelatin type B are used, the molar ratio is 2:1 to 1:2; and when lowBloom gelatin and alginate are used, the molar ratio is 3:1 to 8:1.

Processing aids can be included in the shell material (e.g., primary orouter shells). Processing aids can be used for a variety of reasons. Forexample, they may be used to promote agglomeration of the primarymicrocapsules, stabilize the emulsion system, improve the properties ofthe outer shells, control microcapsule size and/or to act as anantioxidant. In one aspect, the processing aid can be an emulsifier, afatty acid, a lipid, a wax, a microbial cell (e.g., yeast cell lines), aclay, or an inorganic compound (e.g., calcium carbonate). Not wishing tobe bound by theory, these processing aids can improve the barrierproperties of the microcapsules. In one aspect, one or more antioxidantscan be added to the shell material. Antioxidant properties are usefulboth during the process (e.g. during coacervation and/or spray drying)and in the microcapsules after they are formed (i.e. to extendshelf-life, etc). Preferably a small number of processing aids thatperform a large number of functions can be used. In one aspect, theantioxidant can be a phenolic compound, a plant extract, or asulphur-containing amino acid. In one aspect, ascorbic acid (or a saltthereof such as sodium or potassium ascorbate) can be used to promoteagglomeration of the primary microcapsules, to control microcapsule sizeand to act as an antioxidant. The antioxidant can be used in an amountof about 100 ppm to about 12,000 ppm, or from about 1,000 ppm to about5,000 ppm. Other processing aids such as for example, metal chelators,can be used as well. For example, ethylene diamine tetraacetic acid canbe used to bind metal ions, which can reduce the catalytic oxidation ofthe loading substance.

In one aspect, the primary microcapsules (primary shells) have anaverage diameter of about 40 nm to about 10 μm, 1 μm to about 10 μm, 1μm to about 10 μm, 1 μm to about 8 μm, 1 μm to about 6 μm, 1 μm to about4 μm, or 1 μm to about 2 μm, or 1 μm. In another aspect, the multicoremicrocapsules can have an average diameter of from about 1 μm to about2000 μm, 20 μm to about 1000 μm, from about 20 μm to about 100 μm, orfrom about 30 μm to about 80 μm. In another aspect, the single-coremicrocapsules have an outer diameter of from 1 μm to 2,000 μm.

The microcapsules described herein generally have a combination of highpayload and structural strength. For example, payloads of loadingsubstance can be from 20% to 90%, 50% to 70% by weight, or 60% by weightof the single or multicore microcapsules.

In one aspect, the methods disclosed in U.S. Patent ApplicationPublication NO. 2003/0193102, which is incorporated by reference in itsentirety, can be used to encapsulate the disclosed compositions. It isalso contemplated that one or more additional shell layers can be placedon the outer shell of the single or multicore microcapsules. In oneaspect, the techniques described in International Publication No. WO2004/041251 A1, which is incorporated by reference in its entirety, canbe used to add additional shell layers to the single and multicoremicrocapsules.

Targeted Delivery

The disclosed compositions can be targeted to a particular cell type,such as islets cells, via antibodies, receptors, or receptor ligands.The following references are examples of the use of this technology totarget specific tissue (Senter et al., Bioconjugate Chem. 1991,2:447-51; Bagshawe, Br. J. Cancer 1989, 60:275-81; Bagshawe et al., Br.J. Cancer 1988, 58:700-3; Senter et al., Bioconjugate Chem 1993, 4:3-9;Battelli et al., Cancer Immunol. Immunother. 1992, 35:421-5; Pieterszand McKenzie, Immunolog. Rev. 1992, 129:57-80; Roffler et al., Biochem.Pharmacol. 1991, 42:2062-5). These techniques can be used for a varietyof other specific cell types.

Foodstuffs

Also disclosed herein are foodstuffs comprising any of the disclosedcompositions. By “foodstuff” is meant any article that can be consumed(e.g., eaten, drank, or ingested) by a subject. In one aspect, themicrocapsules can be used as nutritional supplements to a foodstuff. Forexample, the microcapsules and emulsions can be loaded with vitamins,omega-3 fatty acids, and other compounds that provide health benefits.In one aspect, the foodstuff is a baked good, a pasta, a meat product, afrozen dairy product, a milk product, a cheese product, an egg product,a condiment, a soup mix, a snack food, a nut product, a plant proteinproduct, a hard candy, a soft candy, a poultry product, a processedfruit juice, a granulated sugar (e.g., white or brown), a sauce, agravy, a syrup, a nutritional bar, a beverage, a dry beverage powder, ajam or jelly, a fish product, or pct companion food. In another aspect,the foodstuff is bread, tortillas, cereal, sausage, chicken, ice cream,yogurt, milk, salad dressing, rice bran, fruit juice, a dry beveragepowder, rolls, cookies, crackers, fruit pies, or cakes.

Methods of Use

The disclosed compositions also have a wide variety of uses. Forexample, the disclosed compositions (including the nutritionalsupplements, pharmaceutical formulations, delivery devices, andfoodstuffs) can be used as a source of fatty acids (e.g., PUFA's likeomega-3 fatty acids), lowering triglycerides and influencing diabetesrelated biochemistry.

In one particular example, disclosed herein are methods of supplementingomega-3 fatty acids in a subject by administering an effective amount ofa composition disclosed herein, wherein the composition comprises anomega-3 fatty acid. In another example, disclosed herein are methods oflowering cholesterol levels, triglyceride levels, or a combinationthereof in a subject by administering an effective amount of acomposition disclosed herein.

In the disclosed methods, the compositions can be any of thecompositions disclosed herein. Also the disclosed compositions can beused neat or in combination with some other component. For example, thedisclosed compositions can be used in the disclosed methods in the formof any of the nutritional supplements disclosed herein. In anotherexample, the disclosed compositions can be used in the disclosed methodsin the form of any of the pharmaceutical formulations disclosed herein.In still another example, the disclosed compositions can be incorporatedin any of the delivery devices disclosed herein, or incorporated intoany foodstuff disclosed herein and used in the disclosed methods.

It is contemplated that the methods disclosed herein can be accomplishedby administering various forms of the disclosed compositions. Forexample, one can administer any of the pharmaceutical formulations withany of the foodstuffs disclosed herein. In another example, one canadminister a microcapsule with any of the nutritional supplementsdisclosed herein. In yet another example, one can administer any of thepharmaceutical formulations with any of the delivery devices andnutritional supplement disclosed herein, and the like.

Dosage

When used in the above described methods or other treatments, or in thenutritional supplements, pharmaceutical formulations, delivery devices,or foodstuffs disclosed herein, an “effective amount” of one of thedisclosed compositions can be employed in pure form or, where such formsexist, in pharmaceutically acceptable salt form, and with or without apharmaceutically acceptable excipient, carrier, or other additive.

The specific effective dose level for any particular subject will dependupon a variety of factors including the disorder being treated and theseverity of the disorder; the identity and activity of the specificcomposition employed; the age, body weight, general health, sex and dietof the patient; the time of administration; the route of administration;the rate of excretion of the specific composition employed; the durationof the treatment; drugs used in combination or coincidental with thespecific composition employed and like factors well known in the medicalarts. For example, it is well within the skill of the art to start dosesof the composition at levels lower than those required to achieve thedesired therapeutic effect and to gradually increase the dosage untilthe desired effect is achieved. If desired, the effective daily dose canbe divided into multiple doses for purposes of administration.Consequently, single dose compositions can contain such amounts orsubmultiples thereof to make up the daily dose.

The dosage can be adjusted by the individual physician or the subject inthe event of any counterindications. Dosage can vary, and can beadministered, in one or more dose administrations daily, for one orseveral days. Guidance can be found in the literature for appropriatedosages for given classes of pharmaceutical products.

Administration and Delivery

In one aspect, disclosed herein are uses of a delivery device to delivera disclosed compositions to a subject. Further, disclosed are methodsfor delivering a disclosed compositions to a subject by administering tothe subject any of the nutritional supplements, pharmaceuticalformulations, delivery devices, and/or foodstuffs disclosed herein.

The disclosed compositions (including nutritional supplements,microcapsules, delivery devices, and pharmaceutical formulations) can beadministered orally, parenterally (e.g., intravenously), byintramuscular injection, by intraperitoneal injection, transdermally,extracorporcally, topically or the like, including topical intranasaladministration or administration by inhalant. As used herein, “topicalintranasal administration” means delivery of the compositions into thenose and nasal passages through one or both of the nares and cancomprise delivery by a spraying mechanism or droplet mechanism, orthrough aerosolization of the nucleic acid or vector. Administration ofthe compositions by inhalant can be through the nose or mouth viadelivery by a spraying or droplet mechanism. Delivery can also bedirectly to any area of the respiratory system (e.g., lungs) viaintubation.

EXAMPLES

The following examples are set forth below to illustrate the methods andresults according to the disclosed subject matter. These examples arenot intended to be inclusive of all aspects of the subject matterdisclosed herein, but rather to illustrate representative methods andresults. These examples are not intended to exclude equivalents andvariations of the present invention which are apparent to one skilled inthe art.

Efforts have been made to ensure accuracy with respect to numbers (e.g.,amounts, temperature, etc.) but some errors and deviations should beaccounted for. Unless indicated otherwise, parts are parts by weight,temperature is in ° C. or is at ambient temperature, and pressure is ator near atmospheric. There are numerous variations and combinations ofconditions, e.g., component concentrations, temperatures, pressures, andother reaction ranges and conditions that can be used to optimize theproduct purity and yield obtained from the described process. Onlyreasonable and routine experimentation will be required to optimize suchprocess conditions.

Certain materials, compounds, compositions, and components disclosedherein can be obtained commercially or readily synthesized usingtechniques generally known to those of skill in the art. For example,the starting materials and reagents used in preparing the disclosedcompositions are either available from commercial suppliers such asOcean Nutrition Canada, Ltd. (Dartmouth, Canada), Aldrich Chemical Co.,(Milwaukee, Wis.), Acros Organics (Morris Plains, N.J.), FisherScientific (Pittsburgh, Pa.), or Sigma (St. Louis, Mo.) or are preparedby methods known to those skilled in the art following procedures setforth in references such as Fieser and Fieser's Reagents for OrganicSynthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry ofCarbon Compounds, Volumes 1-5 and Supplementals (Elsevier SciencePublishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons,1991); March's Advanced Organic Chemistry, (John Wiley and Sons, 4thEdition); and Larock's Comprehensive Organic Transformations (VCHPublishers Inc., 1989).

Example 1

The following examples use TRISYL™, a TRISYL™/clay combination, and clayalone to remove cholesterol in TG 18/12. TG 18/12 is a triglyceridemarine oil with about 18 wt. % EPA and about 12 wt. % DHA. Theseexamples use crude TG 18/12, lot #3929, with a cholesterol, level of 6.5mg/g as the starting oil. Different crude TG 18/12 oils can havedifferent cholesterol contents. The cholesterol removal conditionsdisclosed herein can be adjusted for different oils. For example, crudeTG 18/12, lot #2755, has 4.3 mg/g cholesterol, and thus a mildertreatment can be used to reduce cholesterol to below 1 mg/g.

Adsorbents

TRISYL™ was first tested for cholesterol removal at 80° C. and showed abetter effect than that of clay. However, TRISYL™ does not remove colorwell. A TRISYL™/clay combination was tested for cholesterol removal aswell as bleaching. The amounts of TRISYL™/clay (1:1) in marine oil were10% or above (up to 20%) at 180° C. and the cholesterol level could bereduced to below 1 mg/g (e.g., 0.15 mg/g). Using from 5 to 6%TRISYL™/clay, cholesterol was reduced to from 1.0 to 2.0 mg/g at 180° C.

At high temperatures, clay showed a significantly increased effect toreduce cholesterol. At 80° C., 3% clay reduced about 15% cholesterol inTG 18/12 (lot #3929), while at 180° C., 3% clay removed more than 60%cholesterol in the oil. Using 6% clay in the fish oil at 190° C. for10-20 minutes reduced cholesterol to from 0.5 to 0.8 mg/g. It seemed10-minute treatment was enough to produce cholesterol free oil, althoughmost of the treatments were tested for 20 minutes. For milderconditions, treatment time can be increased to achieve cholesterol freecompositions.

Color

The high temperature treatment under vacuum did not darken the oilcolor. The color of the all after the treatment was lighter than thatafter a normal bleaching procedure (3% clay at 80° C.). While notwishing to be bound by theory, it is believed that one reason for theimproved color is that a higher amount of clay is used in thede-cholesterol procedure.

p-Anisidine Value

Crude TG18/12 (lot #3929) oil had a p-Anisidine value of 20. A normalbleaching with 3% clay and 80° C. reduced the value to 12. Oil samplestreated at high temperature with clay and TRISYL™ had lower p-Anisidinevalues (e.g., from 3 to 7) (see Table 4).

TABLE 4 Effect of treatments on p-Anisidine values and cholesterolcontents Sample information Cholesterol (mg/g) p-Anisidine TG18/12, lot#3929, crude oil (herein “3929 Crude Oil”) 6.55 20.46 Clay:3929 CrudeOil 3:100, 80° C., 60 min 5.45 12.44 Clay:Trisyl:3929 Crude Oil 7:7:100,180° C., 20 min, 0.68 3.89 Clay:Trisyl:3929 Crude Oil 1:1:10, 180° C.,20 min 0.15 4.19 Clay:Trisyl:3929 Crude Oil 6:6:100, 180° C., 20 min0.91 3.60 Clay:3929 Crude Oil 7:100, 180° C., 20 min 0.69 5.42 Clay:3929Crude Oil 6:100, 190° C., 20 min 0.76 5.82 Clay:3929 Crude Oil 5:100,200° C., 20 min 1.16 6.64 Clay:3929 Crude rude Oil 6:100, 200° C., 10min 0.45 5.98 Clay:3929 Crude Oil 6:100, 210° C., 20 min 0.18 7.14Clay:3929 Crude Oil 6:100, 190-200° C., 20 min 0.45 6.93

EPA and DHA

High temperature can destroy EPA and DHA, especially in the presence ofabsorbents. As the temperature and time increased, the loss of EPA andDMA increased (see Table 5). There are about 6 to 7 wt. % EPA and 8 to 9wt. % DHA losses in the cholesterol free oil (e.g., <1 mg/g) using thedisclosed methods. To minimize losses of EPA and DHA, a shortertreatment time can be used.

TABLE 5 Effect of treatments on EPA, DHA, and cholesterol contents EPADHA Sample information Cholesterol (mg/g) (mg/g) (mg/g) 3929 Crude Oil6.55 168.48 117.85 Clay:3929 Crude Oil 7:100, 180° C., 20 min 0.69157.07 108.29 Clay:3929 Crude Oil 6:100, 190° C., 20 min 0.76 157.38108.01 Clay:3929 Crude Oil 5:100, 200° C., 20 min 1.16 157.09 108.45Clay:3929 Crude Oil 6:100, 200° C., 10 min 0.45 153.85 105.10 Clay:3929Crude Oil 6:100, 190-200° C., 20 min 0.45 152.49 103.07 Clay:3929 CrudeOil 6:100, 210° C., 20 min 0.18 123.39 78.97

Lipid Class

The starting TG 18/12, lot #3929, oil has over 98 wt. % of triglyceridesand small amount of diglycerides. After the disclosed treatments, nobreakdown of triglycerides occurred. The diglyceride content was reducedby the treatments. TRISYL™ and clay can absorb compounds (e.g., polarlipids) as well as impurities.

TABLE 6 Effect of treatments on tri-, di-, and mono-glyceride contents(TG, DG, and MG, respectively) Sample information TG (%) DG (%) MG (%)3929 Crude Oil 98.6 1.4 0.0 Clay:3929 Crude Oil 1:1:10, 180° C., 20 min100 0.0 0.0 Clay:3929 Crude Oil 6:100, 200° C., 10 min 99.0 1.0 0.0Clay:3929 Crude Oil 6:100, 190-200° C., 99.2 0.8 0.0 20 min

TABLE 7 Cholesterol removal effects of clay and TRISYL ™ CholesterolSample information (mg/g) TG18/12, lot #4381, crude oil (herein “4381Crude Oil”) 4.25 Trisyl:4381 Crude Oil 1:10, 80° C., 60 min 2.81Clay:4381 Crude Oil 1:1:10, 80° C., 50 min, 180° C., 10 min 0.73Repeated cholesterol analysis 0.71 3929 Crude Oil 6.34 Clay:Trisyl:3929Crude Oil 1:1:10, 180° C., 20 min 0.71 Repeated cholesterol analysis0.70 4381 Crude Oil 4.30 Trisyl:4381 Crude Oil 1:10, 180° C., 20 min,(>80° C. for ~45 min) 1.88 Clay:Trisyl:4381 Crude Oil 1:1:10, 180° C.,20 min, (>80° C. for ~45 min) 0.21 Clay:4381 Crude Oil 1:10, 180 C, 20min, (>80° C. for ~45 min) 0.09 Repeated analysis 0.08 Clay:Trisyl:3604Crude Oil 1:1:10, 180° C., 20 min, (>80° C. for ~45 min) 0.84Trisyl:3604 Crude Oil 1:20, 180° C., 40 min, (>80° C. for ~45 min) 1.883929 Crude Oil 6.50 Clay:Trisyl:3929 Crude Oil 1:1:20, 180° C., 20 min,(>80° C. for ~45 min) 1.80 Clay:Trisyl:3929 Crude Oil 1:1:10, 130° C.,20 min, (>80° C. for ~45 min) 2.44 Clay:Trisyl:3929 Crude Oil 3:3:100,180° C., 20 min, (>80° C. for ~45 min) 1.00 3929 Crude Oil, bottle A6.55 Clay:Trisyl:3929 Crude Oil (bottle A) 3:3:100, 180° C., 20 min,(>80° C. 1.89 for ~45 min) Clay:Trisyl:3929 Crude Oil (bottle A) 1:1:20,130° C., 40 min, (>80° C. for 2.52 ~60 min) Clay:3929 Crude Oil (bottleA) 1:20, 180° C., 20 min, (>80° C. for ~50 min) 1.22 Clay:Trisyl:3929Crude Oil (bottle A) oil 7:7:100, 180° C., 20 min, (>80° C. 0.68 for ~45min) Clay:Trisyl:3929 Crude Oil (bottle A) 1:1:10, 160° C., 20 min,(>80° C. for 1.18 ~45 min) Clay:Trisyl:3929 Crude Oil (bottle A) 1:1:10,180° C., 20 min, (>80° C. for 0.15 ~45 min) Clay:Trisyl:3929 Crude Oil(bottle A) 6:6:100, 180° C., 20 min, (>80° C. 0.91 for ~45 min)Clay:Trisyl:3929 Crude Oil (bottle A) 1:1:10, 180° C., 10 min, (>80° C.for 0.59 ~35 min) Clay:Trisyl:3929 Crude Oil (bottle A) 1:1:20, 160° C.,20 min, (>80° C. for 2.04 ~45 min) Clay:Trisyl:3929 Crude Oil (bottle A)3:3:100, 180° C., 20 min, (>80° C. 1.07 for ~45 min) Clay:Trisyl:3929Crude Oil (bottle A) 1:1:10, 180° C., 20 min, (>80° C. for 0.34 ~45 min)Clay:Trisyl:3929 Crude Oil (bottle A) 1:1:20, 180° C., 20 min, (>80° C.for 0.74 ~45 min) 3929 Crude Oil 6.40 Clay:Trisyl:3929 Crude Oil2:3:100, 180° C., 20 min, (>80° C. for ~45 min) 2.11 Clay:Trisyl:3929Crude Oil (A) 2:3:100, 180° C., 20 min, (>80° C. for ~45 min) 1.43Clay:Trisyl:3929 Crude Oil 1:1:40, 180° C., 20 min, (>80° C. for ~45min) 1.50 Clay:Trisyl:oil from sample immediately above 1:1:40, 180° C.,20 min, 1.06 (>80° C. for ~45 min) Clay:Trisyl:3929 Crude Oil 2:3:100,160° C., 20 min, (>80° C. for ~45 min) 2.32 Clay:Trisyl:oil from sampleimmediately above 2:3:100, 160° C. 20 min, 1.93 (>80° C. for ~45 min)Clay:3929 Crude Oil 6:100, 180° C., 20 min, (>80° C. for ~45 min) 1.19Clay:3929 Crude Oil 6:100, 160° C., 20 min, (>80° C. for ~45 min) 1.69Clay:3929 Crude Oil 3:100, 180° C., 20 min, (>80° C. for ~45 min) 2.33Clay:oil from sample immediately above 3:100, 180° C., 20 min, (>80° C.1.51 for ~45 min) Clay:3929 Crude Oil 3:100, 80° C., 60 min, 5.45Clay:3929 Crude Oil 7:100, 180° C., 20 min, (>80° C. for ~45 min) 0.69Clay:3929 Crude Oil 7:100, 170° C., 20 min, (>80° C. for ~45 min) 1.37Clay:3929 Crude Oil 5:100, 190° C., 20 min, (>80° C. for ~45 min) 1.07Clay:3929 Crude Oil 6:100, 190° C., 20 min, (>80° C. for ~45 min) 0.76Clay:3929 Crude Oil 5:100, 200° C., 20 min, (>80° C. for ~45 min) 1.16Clay:3929 Crude Oil 5:100, 190° C., 20 min, (>80° C. for ~45 min) 1.15Clay:3929 Crude Oil 6:100, 200° C., 10 min, (>80° C. for ~35 min) 0.45Clay:3929 Crude Oil 6:100, 210° C., 20 min, (>80° C. for ~45 min) 0.18Clay:3929 Crude Oil 6:100, 190-200° C., 20 min, (>80° C. for ~45 min)0.45 TG18/12, lot #2755, crude oil (herein “2755 Crude Oil”) 4.31Clay:2755 Crude Oil 6:100, 180° C., 20 min, (>80° C. for ~45 min) 0.73

Results

At medium temperature (about 80° C.), TRISYL™ showed a better effect toremove cholesterol from the oil than that of clay. The combination ofTRISYL™ and clay reduced up to 56 wt. % of cholesterol in the oil butdid not reduce the cholesterol level below 2 mg/g. At highertemperatures (e.g., greater than about 180° C.), clay showed asignificantly increased effect to remove cholesterol. Clay alone or acombination of TRISYL™/clay were found to be able to remove up to 98 wt.% of cholesterol. The conditions of using 5 wt. % clay at 180 to 190° C.were found to be able to reduce cholesterol levels to 1.0 to 1.5 mg/g.The conditions of using 6 to 7 wt. % clay at 180 to 190° C. were foundto be able to reduce cholesterol level to below 1.0 mg/g.

About 6.5% EPA and about 8.5% DHA were lost using these methods. Alsode-cholesterol samples had lower p-Anisidine values as compared to thatof the starting oil. Further, lipid class analysis showed that thetreatments reduced mono- and di-glycerides.

Example 2

Seven TG 18/12 oils were tested for milder conditions. They were crudeoils with lot numbers 43929, #2755 and #3985, and steam deodorized oilswith lot numbers 45147, #4254 (two bottles with different cholesterolcontents), and #5139. These oils are identified respectively herein as3929 Crude Oil, 2755 Crude Oil, 3985 Crude Oil, 5147 Crude Oil, 4254Crude Oil (Bottle A and Bottle B), and 5139 Crude Oil.

The temperatures were chosen from 140 to 160° C. based on the previousexperiment results. Temperature can be a factor to remove cholesterol infish oil using clay. Table 8 shows how temperatures affect cholesterolremoval with 6% clay.

TABLE 8 Effect of temperatures on cholesterol removal Cholesterol Sampleinformation (mg/g) 3929 Crude Oil 6.40 Clay:3929 Crude Oil 6:100, 160°C., 20 min 1.69 Clay:3929 Crude Oil 6:100, 180° C., 20 min 1.19Clay:3929 Crude Oil 6:100, 190° C., 20 min 0.76 Clay:3929 Crude Oil6:100, 200° C., 10 min 0.45 Clay:3929 Crude Oil 6:100, 210° C., 20 min0.18

Table 9 shows the results at 140° C. for 20 min. The cholesterol levelswere slightly above 2 mg/g after the treatments. For normal fish oils(with cholesterol level between 4-7 mg/g), the cholesterol contentsafter the treatment were most likely to be 2.0 to 2.5 mg/g. In order toreduce cholesterol level below 2 mg/g, increase of treatment time ortemperature can be used.

TABLE 9 Cholesterol removal at about 140° C. Cholesterol Sampleinformation (mg/g) 2755 Crude Oil 5.43 Clay:2755 Crude Oil 6:100, 140°C., 20 min 2.16 Clay:2755 Crude Oil 6:100, 140° C., 20 min 2.25Clay:2755 Crude Oil 5:100, 140-150° C., 20 min 2.14 Clay:2755 Crude Oil5:100, 140° C., 20 min 2.25 Clay:2755 Crude Oil 6:100, 150° C., 20 min2.05 Clay:2755 Crude Oil 6:100, 150° C., 20 min 2.10

After treatment at 160° C., the cholesterol contents in most of the fishoil tested (six of the seven oils) were below 2 mg/g (see Table 10). Toprevent damage of fish oil, a 20 minute treatment time was chosen. Forrelatively mild conditions (140 to 160° C.), the treatment time could beincreased to 40 to 60 minutes, which can further reduce cholesterollevels.

TABLE 10 Cholesterol removal at 160° C. with 6% clay Cholesterol Sampleinformation (mg/g) 3929 Crude Oil 6.40 Clay:3929 Crude Oil 6:100, 160°C., 20 min 1.69 2755 Crude Oil 5.43 Clay:2755 Crude Oil 6:100, 160° C.,20 min 1.96 5147 Crude Oil (which is steam deodorized) 5.25 Clay:5147Crude Oil 6:100, 160 C, 20 min 1.97 4254 Crude Oil Bottle A (which issteam deodorized) 5.06 Clay:4254 Crude Oil Bottle A 6:100, 160° C., 20min 1.91 4254 Crude Oil Bottle B (which is steam deodorized) 5.55Clay:4254 Crude Oil Bottle B 6:100, 160° C., 20 min 1.96 5139 Crude Oil(which is steam deodorized) 5.18 Clay:5139 Crude Oil 6:100, 160° C., 20min 1.98 3985 Crude Oil 7.11 Clay:3985 Crude Oil 6:100, 160° C., 20 min2.89

The disclosed methods also served as bleaching procedures which producedlighter oil compared to that of normal bleaching procedure. Hightemperature bleaching did not show negative effects on the fish oilcolor.

Trans Fatty Acid

Trans-fatty acid contents in the fish oils before and afterde-cholesterol procedure were tested. The oils were transferred intofatty acid methyl esters (FAMEs) and analyzed for trans-fatty acids.Ethyl EPA standard (all cis-EPA) was also transesterified into its FAME,to monitor how much trans-fatty acid might be produced during thetransesterification. The results (Table 11) are based on area percentage(not quantified).

TABLE 11 Trans fatty acids in fish oils before and after treatment TransTrans EPA Trans Trans DHA Sample EPA (rep.) DHA (rep.) 3985 Crude Oil1.2 1.2 1.1 1.5 After 160° C., 6% clay 1.2 1.4 1.4 1.5 treatment 5147Crude Oil (which is 5.7 5.1 5.9 5.8 steam deodorized) After 160 C, 6%clay 7.4 5.9 7.6 7.1 treatment

Crude TG 18/12 oil, lot #3985, does not have much trans-fatty acids,while steam deodorized oil has a higher amount of trans-fatty acids. Thedisclosed procedures can be used before Steam deodorization.

For the crude oil, the disclosed procedures did not greatly increasetrans-fatty acids. But for the steam deodorized oil, after the disclosedtreatments, trans-fatty acids increase significantly. While not wishingto be bound by theory, it is believed that the steam deodorizationproduce itself increases trans-fatty acids and makes the oil vulnerableto the following treatments.

TABLE 12 Cholesterol removal results with clay Cholesterol Sampleinformation (mg/g) 3929 Crude Oil 6.40 Clay:3929 Crude Oil 6:100, 160°C., 20 min 1.69 Clay:3929 Crude Oil 6:100, 180° C., 20 min 1.19Clay:3929 Crude Oil 6:100, 190° C., 20 min 0.76 Clay:3929 Crude Oil6:100, 200° C., 10 min 0.45 Clay:3929 Crude Oil 6:100, 210° C., 20 min0.18 2755 Crude Oil 5.43 Clay:2755 Crude Oil 6:100, 140° C., 20 min 2.16Clay:2755 Crude Oil 6:100, 140° C., 20 min 2.25 Clay:2755 Crude Oil6:100, 150° C., 20 min 2.05 Clay:2755 Crude Oil 6:100, 160° C., 20 min1.96 Clay:2755 Crude Oil 6:100, 180° C., 20 min 0.73 3985 Crude Oil,Bottle A 7.15 3985 Crude Oil, Bottle B 7.11 Clay:3985 Crude Oil Bottle A6:100, 140° C., 20 min 3.22 Clay:3985 Crude Oil Bottle A 6:100, 140° C.,20 min 3.19 Clay:3985 Crude Oil Bottle A 6:100, 160° C., 20 min 2.89Clay:3985 Crude Oil Bottle B 6:100, 170° C., 20 min 2.61 Clay:3985 CrudeOil Bottle A 6:100, 170° C., 20 min 2.34 5147 Crude Oil (which is steamdeodorized) 5.25 5147 Crude Oil repeat 5.35 Clay:5147 Crude Oil 6:100,160° C., 20 min 1.97 Clay:5147 Crude Oil 5:100, 160° C., 20 min 2.104254 Crude Oil Bottle A (which is steam deodorized) 5.06 Clay:4254 CrudeOil Bottle A 5:100, 160° C., 20 min 1.95 Clay:4254 Crude Oil Bottle A6:100, 160° C., 20 min 1.91 4254 Crude Oil Bottle B (which is steamdeodorized) 5.55 Clay:4254 Crude Oil Bottle B 6:100, 160° C., 20 min1.96 Clay:4254 Crude Oil Bottle B 5:100, 160° C., 20 min 2.00 5139 CrudeOil (which is steam deodorized) 5.18 Clay:5139 Crude Oil 6:100, 160° C.,20 min 1.98 Clay:5139 Crude Oil 5:100, 160° C., 20 min 2.06

Example 3

The effects of high-temperature-bleaching on oil qualities, cholesterolreduction, changes of EPA/DHA ratios and amounts, changes in trans-fattyacids, color, and oxidative stability were tested. Oxidative stabilitycan be measured by well known methods, such as the Active Oxygen Method(AOM) and Oxygen Stability Index (OSI). These methods measure the extentof oxidation to provide an indication of the point at which oxidativerancidity occurs. Suitable methods form measuring oxidative stabilitycan involve the use of the Oxidation Stability Instrument (availablefrom Omnion Inc., Rockland, Mass.) and the Rancimat (available fromBrinkman Instruments, Des Plaines, Ill.). These instruments measure thechanges in conductivity caused by ionic volatile organic acids. The endpoint is selected as the point at which the rapid rise in conductancebegins (i.e., the oxidative stability induction time).

TG oil, lot #8823, which was alkaline refined, was obtained fromMulgrave and Englehard clay. The processing temperature was at 150° C.60 g of oil was mixed with 3 g clay. The mixture was placed undervacuum, stirred, and heated to 150° C. The mixture was allowed tocontinue stirring at 150° C. for 30 minutes. After cooling to 50° C.(about 40 min), the vacuum was broken with nitrogen. The mixture wasthen filtered through filtration paper.

TABLE 13 Effect of temperatures on cholesterol removal and oil qualitiesOxidative stability Cholesterol induction Trans Color Sample processing(mg/g) time (h) EPA/DHA (wt. %) (Gardner) 8823 Crude Oil 7.05 0.65158/139 11.5 yellow/brown 30 min at 140° C. 1.79 1.40 30 min at 140° C.1.57 1.20 double bleaching each with 5% clay 45 min at 140° C. 1.70 1.131.5 45 min at 140° C. 1.15 1.20 double bleaching each with 5% clay 30min at 150° C. 1.73 1.10 151/142 1.3 30 min at 150° C. 1.74 1.05 1.2 30min at 150° C. 1.23 0.90 double bleaching each with 5% clay 30 min at150° C. 0.83 0.85 144/137 1.8 double bleaching each with 5% clay 45 minat 150° C. 1.47 1.00 150/142 1.2 3.5 very light yellow 45 min at 150° C.1.25 0.90 4.0 30 min at 160° C. 1.70 1.15 1.3 30 min at 160° C. 1.600.75 1.3 20 min at 180° C. 1.28 0.80 2.4 20 min at 180° C. 0.63 0.55 2.4

The starting oil (lot #8822) has a typical cholesterol level of 7 mg/g.The disclosed procedures were able to reduce the level to below 2.0mg/g. The higher or longer the temperature or processing time, the lowerthe cholesterol level. The oxidative stability induction time of theoils treated increased.

It will be apparent to those skilled in the art that variousmodifications and variations can be made in the present inventionwithout departing from the scope or spirit of the invention. Otherembodiments of the invention will be apparent to those skilled in theart from consideration of the specification and practice of theinvention disclosed herein. It is intended that the specification andexamples be considered as exemplary only, with a true scope and spiritof the invention being indicated by the following claims.

1-73. (canceled)
 74. A composition comprising fish oil, wherein thecomposition comprises less than about 2 mg of sterol per gram ofcomposition, at least about 97% triglycerides, less than about 2%diglycerides, and less than about 1% monoglycerides.
 75. The compositionof claim 74, wherein the fish oil comprises an Atlantic fish oil,Pacific fish oil, Mediterranean fish oil, light pressed fish oil,alkaline treated fish oil, heat treated fish oil, light and heavy brownfish oil, tuna oil, bonito oil, sea bass oil, halibut oil, spearfishoil, barracuda oil, cod oil, menhaden oil, sardine oil, pilchard oil,anchovy oil, capelin oil, Atlantic cod oil, Atlantic herring oil,Atlantic mackerel oil, Atlantic menhaden oil, salmonids oil, orcombination thereof.
 76. The composition of claim 74, wherein thecomposition comprises an EPA to DHA wt. % ratio of about 60:0.3, about18:12, about 5:25, or about 0.8:60.
 77. The composition of claim 74,wherein the composition comprises from about 1.1 to about 0.9 milligramsof sterol per gram of the composition.
 78. The composition of claim 74,wherein the composition comprises about 1.0 milligram of sterol per gramof the composition.
 79. The composition of claim 74, wherein the sterolis cholesterol.
 80. The composition of claim 79, wherein the cholesterolis in its free form and esterified form.
 81. The composition of claim74, wherein the composition has a Gardner color of less than or equal toabout 5, as determined by ASTM D
 1544. 82. The composition of claim 74,wherein the composition comprises linolenic acid, octadecatetraenoicacid, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA),docosapentaenoic acid (DPA), or a residue, derivatives, or mixturethereof.
 83. The composition of claim 74, wherein the compositioncomprises from about 10 to about 16 wt. % DHA.
 84. The composition ofclaim 74, wherein the composition comprises from about 14 to about 20wt. % EPA.
 85. The composition of claim 74, wherein the composition hasless than or equal to about 2 wt. % trans-fatty acids.
 86. Thecomposition of claim 74, wherein the composition has a p-Anisidine valueof less than or equal to about 10, as determined by ISO 6885:1998. 87.The composition of claim 74, wherein the composition has a p-Anisidinevalue of less than or equal to about 6, as determined by ISO 6885:1998.